Chemicals Commonly Used in Biochemistry and Molecular Biology and Their Properties

Authored by: Roger L. Lundblad

Biochemistry and Molecular Biology Compendium

Print publication date:  June  2007
Online publication date:  June  2007

Print ISBN: 9781420043471
eBook ISBN: 9781420043488
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Tunnicliff, G. and Smith, J.A., Competitive inhibition of gamma-aminobutyric acid receptor binding by N-hydroxyethylpiperazine-N′-2-ethanesulfonic acid and related buffers, J. Neurochem. 36, 1122–1126, 1981; Chappel, D.J., N-[(carbamoylmethyl) amino] ethanesulfonic acid improves phenotyping of α-1-antitrypsin by isoelectric focusing on agarose gel, Clin. Chem. 31, 1384–1386, 1985; Liu, Q., Li, X., and Sommer, S.S., pk-matched running buffers for gel electrophoresis, Anal. Biochem. 270, 112–122, 1999; Taha, M., Buffers for the physiological pH range: acidic dissociation constants of zwitterionic compounds in various hydroorganic media, Ann. Chim. 95, 105–109, 2005.

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Chemicals Commonly Used in Biochemistry and Molecular Biology and Their Properties


Tunnicliff, G. and Smith, J.A., Competitive inhibition of gamma-aminobutyric acid receptor binding by N-hydroxyethylpiperazine-N′-2-ethanesulfonic acid and related buffers, J. Neurochem. 36, 1122–1126, 1981; Chappel, D.J., N-[(carbamoylmethyl) amino] ethanesulfonic acid improves phenotyping of α-1-antitrypsin by isoelectric focusing on agarose gel, Clin. Chem. 31, 1384–1386, 1985; Liu, Q., Li, X., and Sommer, S.S., pk-matched running buffers for gel electrophoresis, Anal. Biochem. 270, 112–122, 1999; Taha, M., Buffers for the physiological pH range: acidic dissociation constants of zwitterionic compounds in various hydroorganic media, Ann. Chim. 95, 105–109, 2005.


Burton, R.M. and Stadtman, E.R., The oxidation of acetaldehyde to acetyl coenzyme A, J. Biol. Chem. 202, 873–890, 1953; Gruber, M. and Wesselius, J.C., Nature of the inhibition of yeast carboxylase by acetaldehyde, Biochim. Biophys. Acta 57, 171–173, 1962; Holzer, H., da Fonseca-Wollheim, F., Kohlhaw, G., and Woenckhaus, C.W., Active forms of acetaldehyde, pyruvate, and glycolic aldehyde, Ann. N.Y. Acad. Sci. 98, 453–465, 1962; Brooks, P.J. and Theruvathu, J.A., DNA adducts from acetaldehyde: implications for alcohol-related carcinogenesis, Alcohol 35, 187–193, 2005; Tyulina, O.V., Prokopieva, V.D., Boldyrev, A.A., and Johnson, P., Erthyrocyte and plasma protein modification in alcoholism: a possible role of acetaldehyde, Biochim. Biophys. Acta 1762, 558–563, 2006; Pluskota-Karwatka, D., Pawlowicz, A.J., and Kronberg, L., Formation of malonaldehyde-acetaldehyde conjugate adducts in calf thymus DNA, Chem. Res. Toxicol. 19, 921–926, 2006.


Banfield, A.G., Age changes in the acetic acid-soluble collagen in human skin, Arch. Pathol. 68, 680–684, 1959; Steven, F.S. and Tristram, G.R., The denaturation of acetic acid-soluble calf-skin collagen. Changes in optical rotation, viscosity, and susceptibility towards enzymes during serial denaturation in solutions of urea, Biochem. J. 85, 207–210, 1962; Neumark, T. and Marot, I., The formation of acetic-acid soluble collagen under polarization and electron microscrope, Acta Histochem. 23, 71–79, 1966; Valfleteren, J.R., Sequential two-dimensional and acetic acid/urea/Triton X-100 gel electrophoresis of proteins, Anal. Biochem. 177, 388–391, 1989; Smith, B.J., Acetic acid-urea polyacrylamide gel electrophoresis of proteins, Methods Mol.Biol. 32, 39–47, 1994; Banfield, W.G., MacKay, C.M., and Brindley, D.C., Quantitative changes in acetic acid-extractable collagen of hamster skin related to anatomical site and age, Gerontologia 12, 231–236, 1996; Lian, J.B., Morris, S., Faris, B. et al., The effects of acetic acid and pepsin on the crosslinkages and ultrastructure of corneal collagen, Biochim. Biophys. Acta. 328, 193–204, 1973; Canto, M.I., Chromoendoscopy and magnifying endoscopy for Barrett’s esophagus, Clin.Gastroenterol.Hepatol. 3 (7 Suppl. 1), S12–S15, 2005; Sionkowska, A., Flash photolysis and pulse radiolysis studies on collagen Type I in acetic acid solution, J. Photochem. Photobiol. B 84, 38–45, 2006.


Jencks, W.P., Barley, F., Barnett, R., and Gilchrest, M., The free energy of hydrolysis of acetic anhydride, J. Am. Chem. Soc. 88, 4464–4467, 1966; Cromwell, L.D. and Stark, G.D., Determination of the carboxyl termini of proteins with ammonium thiocyanate and acetic anhydride, with direct identification of the thiohydantoins, Biochemistry 8, 4735–4740, 1969; Montelaro, R.C. and Rueckert, R.R., Radiolabeling of proteins and viruses in vitro by acetylation with radioactive acetic anhydride, J. Biol. Chem. 250, 1413–1421, 1975; Valente, A.J. and Walton, K.W., The binding of acetic anhydride- and citraconic anhydride-modified human low-density lipoprotein to mouse peritoneal macrophages. The evidence for separate binding sites, Biochim. Biophys. Acta 792, 16–24, 1984; Fojo, A.T., Reuben, P.M., Whitney, P.L., and Awad, W.M., Jr., Effect of glycerol on protein acetylation by acetic anhydride, Arch. Biochem. Biophys. 240, 43–50, 1985; Buechler, J.A., Vedvick, T.A., and Taylor, S.S., Differential labeling of the catalytic subunit of cAMP-dependent protein kinase with acetic anhydride: substrate-induced conformational changes, Biochemistry 28, 3018–3024, 1989; Baker, G.B., Coutts, R.T., and Holt, A., Derivatization with acetic anhydride: applications to the analysis of biogenic amines and psychiatric drugs by gas chromatography and mass spectrometry, J. Pharmacol. Toxicol. Methods 31, 141–148, 1994; Ohta, H., Ruan, F., Hakomori, S., and Igarashi, Y., Quantification of free Sphingosine in cultured cells by acetylation with radioactive acetic anhydride, Anal. Biochem. 222, 489–494, 1994; Yadav, S.P., Brew, K., and Puett, D., Holoprotein formation of human chorionic gonadotropin: differential trace labeling with acetic anhydride, Mol. Endocrinol. 8, 1547–1558, 1994; Miyazaki, K. and Tsugita, A., C-terminal sequencing method for peptides and proteins by the reaction with a vapor of perfluoric acid in acetic anhydride, Proteomics 4, 11–19, 2004.


La Du, B., Jr. and Greenberg, D.M., The tyrosine oxidation system of liver. I. Extracts of rat liver acetone powder, J. Biol. Chem. 190, 245–255, 1951; Korn, E.D. and Payza, A.N., The degradation of heparin by bacterial enzymes. II. Acetone powder extracts, J. Biol. Chem. 223, 859–864, 1956; Ohtsuki, K., Taguchi, K., Sato, K., and Kawabata, M., Purification of ginger proteases by DEAE-Sepharose and isoelectric focusing, Biochim. Biophys. Acta 1243, 181–184, 1995; Selden, L.A., Kinosian, H.J., Estes, J.E., and Gershman, L.C., Crosslinked dimers with nucleating activity in actin prepared from muscle acetone powder, Biochemistry 39, 64–74, 2000; Abadir, W.F., Nakhla, V., and Chong, F., Removal of superglue from the external ear using acetone: case report and literature review, J. Laryngol. Otol. 109, 1219–1221, 1995; Jones, A.W., Elimination half-life of acetone in humans: case reports and review of the literature, J. Anal. Toxicol. 24, 8–10, 2000; Huang, L.P. and Guo, P., Use of acetone to attain highly active and soluble DNA packaging protein Gp16 of Phi29 for ATPase assay, Virology 312, 449–457, 2003; Paska, C., Bogi, K., Szilak, L. et al., Effect of formalin, acetone, and RNAlater fixatives on tissue preservation and different size amplicons by real-time PCR from paraffin-embedded tissues, Diagn. Mol. Pathol. 13, 234–240, 2004; Kuksis, A., Ravandi, A., and Schneider, M., Covalent binding of acetone to aminophospholipids in vitro and in vivo, Ann. N.Y. Acad. Sci. 1043, 417–439, 2005; Perera, A., Sokolic, F., Almasy, L. et al., On the evaluation of the Kirkwood–Buff integrals of aqueous acetone mixtures, J. Chem. Physics 123, 23503, 2005; Zhou, J., Tao, G., Liu, Q. et al., Equilibrium yields of mono- and di-lauroyl mannoses through lipase-catalyzed condensation in acetone in the presence of molecular sieves, Biotechnol. Lett. 28, 395–400, 2006.


Hodgikinson, S.C. and Lowry, P.J., Hydrophobic-interaction chromatography and anion-exchange chromatography in the presence of acetonitrile. A two-step purification method for human prolactin, Biochem. J. 199, 619–627, 1981; Wolf-Coporda, A., Plavsic, F., and Vrhovac, B., Determination of biological equivalence of two atenolol preparations, Int. J. Clin. Pharmacol. Ther. Toxicol. 25, 567–571, 1987; Fischer, U., Zeitschel, U., and Jakubke, H.D., Chymotrypsin-catalyzed peptide synthesis in an acetonitrile-water-system: studies on the efficiency of nucleophiles, Biomed. Biochim. Acta 50, S131–S135, 1991; Haas, R. and Rosenberry, T.L., Protein denaturation by addition and removal of acetonitrile: application to tryptic digestion of acetylcholinesterase, Anal. Biochem. 224, 425–427, 1995; Joansson, A., Mosbach, K., and Mansson, M.O., Horse liver alcohol dehydrogenase can accept NADP+ as coenzyme in high concentrations of acetonitrile, Eur. J. Biochem. 227, 551–555, 1995; Barbosa, J., Sanz-Nebot, V., and Toro, I., Solvatochromic parameter values and pH in acetonitrile-water mixtures. Optimization of mobile phase for the separation of peptides by high-performance liquid chromatography, J. Chromatog. A 725, 249–260, 1996; Barbosa, J., Hernandez-Cassou, S., Sanz-Nebot, V., and Toro, I., Variation of acidity constants of peptides in acetonitrile-water mixtures with solvent composition: effect of preferential salvation, J. Pept. Res. 50, 14–24, 1997; Badock, V., Steinhusen, U., Bommert, K., and Otto, A., Prefractionation of protein samples for proteome analysis using reversed-phase high-performance liquid chromatography, Electrophoresis 22, 2856–2864, 2001; Yoshida, T., Peptide separation by hydrophilic-interaction chromatography: a review, J. Biochem. Biophys. Methods 60, 265–280, 2004: Kamau, P. and Jordan, R.B., Complex formation constants for the aqueous copper(I)-acetonitrile system by a simple general method, Inorg. Chem. 40, 3879–3883, 2001; Nagy, P.I. and Erhardt, P.W., Monte Carlo simulations of the solution structure of simple alcohols in water-acetonitrile mixtures, J. Phys. Chem. B Condens. Matter Mater. Surf. Interfaces Biophys. 109, 5855–5872, 2005; Kutt, A., Leito, I., Kaljurand, I. et al., A comprehensive self-consistent spectrophotometric acidity scale of neutral Bronstad acids in acetonitrile, J. Org. Chem. 71, 2829–2938, 2006.


Hallaq, Y., Becker, T.C., Manno, C.S., and Laposata, M., Use of acetyl chloride/methanol for assumed selective methylation of plasma nonesterified fatty acids results in significant methylation of esterified fatty acids, Lipids 28, 355–360, 1993; Shenoy, N.R., Shively, J.E., and Bailey, J.M., Studies in C-terminal sequencing: new reagents for the synthesis of peptidylthiohydantoins, J. Protein Chem. 12, 195–205, 1993; Bosscher, G., Meetsma, A., and van De Grampel, J.C., Novel organo-substituted cyclophosphazenes via reaction of a monohydro cyclophosphazene and acetyl chloride, Inorg. Chem. 35, 6646–6650, 1996; Mo, B., Li, J., and Liang, S., A method for preparation of amino acid thiohydantoins from free amino acids activated by acetyl chloride for development of protein C-terminal sequencing, Anal. Biochem. 249, 207–211, 1997; Studer, J., Purdie, N., and Krouse, J.A., Friedel–Crafts acylation as a quality control assay for steroids, Appl. Spectros. 57, 791–796, 2003.


Szasz, G., Gruber, W., and Bernt, E., Creatine kinase in serum. I. Determination of optimum reaction conditions, Clin. Chem. 22, 650–656, 1976; Holdiness, M.R., Clinical pharmacokinetics of N-acetylcysteine, Clin. Pharmacokinet. 20, 123–134, 1991; Kelley, G.S., Clinical applications of N-acetylcysteine, Altern. Med. Rev. 3, 114–127, 1998; Schumann, G., Bonora, R., Ceriotti, F. et al., IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37°C. Part 2. Reference procedure for the measurement of catalytic concentration of creatine kinase, Clin. Chem. Lab. Med. 40, 635–642, 2002; Zafarullah, M., Li, W.Q., Sylvester, J., and Ahmad, M., Molecular mechanisms of N-acetylcysteine actions, Cell. Mol. Life Sci. 60, 6–20, 2003; Marzullo, L., An update of N-acetylcysteine treatment for acute aminoacetophen toxicity in children, Curr. Opin. Pediatr. 17, 239–245, 2005; Aitio, M.L., N-acetylcysteine — passépartout or much ado about nothing? Br. J. Clin. Pharmacol. 61, 5–15, 2006.


Lundblad, R.L., Chemical Reagents for Protein Modification, CRC Press, Boca Raton, FL, 2004; Gorbunoff, M.J., Exposure of tyrosine residues in proteins. 3. The reaction of cyanuric fluoride and N-acetylimidazole with ovalbumin, chymotrypsinogen, and trypsinogen, Biochemistry 44, 719–725, 1969; Houston, L.L. and Walsh, K.A., The transient inactivation of trypsin by mild acetylation with N-acetylimidazole, Biochemistry 9, 156–166, 1970; Shifrin, S. and Solis, B.G., Reaction of N-acetylimidazole with L-asparaginase, Mol. Pharmacol. 8, 561–564, 1972; Ota, Y., Nakamura, H., and Samejima, T., The change of stability and activity of thermolysin by acetylation with N-acetylimidazole, J. Biochem. 72, 521–527, 1972; Kasai, H., Takahashi, K., and Ando, T., Chemical modification of tyrosine residues in ribonuclease T1 with N-acetylimidazole and p-diazobenzenesulfonic acid, J. Biochem. 81, 1751–1758, 1977; Zhao, X., Gorewit, R.C., and Currie, W.B., Effects of N-acetylimidazole on oxytocin binding in bovine mammary tissue, J. Recept. Res. 10, 287–298, 1990; Wells, I. and Marnett, L.J., Acetylation of prostaglandin endoperoxide synthase by N-acetylimidazole: comparison to acetylation by aspirin, Biochemistry 31, 9520–9525, 1992; Cymes, G.D., Iglesias, M.M., and Wolfenstein-Todel, C., Chemical modification of ovine prolactin with N-acetylimidazole, Int. J. Pept. Protein Res. 42, 33–38, 1993; Zhang, F., Gao, J., Weng, J. et al., Structural and functional differences of three groups of tyrosine residues by acetylation of N-acetylimidazole in manganese-stabilizing protein, Biochemistry 44, 719–725, 2005.


Hawkins, D., Pinckard, R.N., and Farr, R.S., Acetylation of human serum albumin by acetylsalicylic acid, Science 160, 780–781, 1968; Kalatzis, E., Reactions of aminoacetophen in pharmaceutical dosage forms: its proposed acetylation by acetylsalicylic acid, J. Pharm. Sci. 59, 193–196, 1970; Pinckard, R.N., Hawkins, D., and Farr, R.S., The inhibitory effect of salicylate on the actylation of human albumin by acetylsalicylic acid, Arthritis Rheum. 13, 361–368, 1970; Van Der Ouderaa, F.J., Buytenhek, M., Nugteren, D.H., and Van Dorp, D.A., Acetylation of prostaglandin endoperoxide synthetase with acetylsalicylic acid, Eur. J. Biochem. 109, 1–8, 1980; Rainsford, K.D., Schweitzer, A., and Brune, K., Distribution of the acetyl compared with the salicyl moiety of acetylsalicylic acid. Acetylation of macromolecules in organs wherein side effects are manifest, Biochem. Pharmacol. 32, 1301–1308, 1983; Liu, L.R. and Parrott, E.L., Solid-state reaction between sulfadiazine and acetylsalicyclic acid, J. Pharm. Sci. 80, 564–566, 1991; Minchin, R.F., Ilett, K.F., Teitel, C.H. et al., Direct O-acetylation of N-hydroxy arylamines by acetylsalicylic acid to form carcinogen-DNA adducts, Carcinogenesis 13, 663–667, 1992.


Eftink, M.R. and Ghiron, C.A., Fluorescence quenching studies with proteins, Anal. Biochem. 114, 199–227, 1981; Dearfield, K.L., Abernathy, C.O., Ottley, M.S. et al., Acrylamide: its metabolism, developmental and reproductive effects, Mutat. Res. 195, 45–77, 1988; Williams, L.R., Staining nucleic acids and proteins in electrophoresis gels, Biotech. Histochem. 76, 127–132, 2001; Hamden, M., Bordini, E., Galvani, M., and Righetti, P.G., Protein alkylation by acrylamide, its N-substituted derivatives and crosslinkers and its relevance to proteomics: a matrix-assisted laser desorption/ionization-time of flight-mass spectrometry study, Electrophoresis 22, 1633–1644, 2001; Cioni, P. and Strambini, G.B., Tryptophan phosphorescence and pressure effects on protein structure, Biochim. Biophys. Acta 1595, 116–130, 2002; Taeymans, D., Wood, J., Ashby, P. et al., A review of acrylamide: an industry perspective on research, analysis, formation, and control, Crit. Rev. Food Sci. Nutr. 44, 323–347, 2004; Rice, J.M., The carcinogenicity of acrylamide, Mutat. Res. 580, 3–20, 2005; Besaratinia, A. and Pfeifer, G.P., DNA adduction and mutagenic properties of acrylamide, Mutat. Res. 580, 31–40, 2005; Hoenicke, K. and Gaterman, R., Studies on the stability of acrylamide in food during storage, J. AOAC Int. 88, 268–273, 2005; Castle, L. and Ericksson, S., Analytical methods used to measure acrylamide concentrations in foods, J. AOAC Int. 88, 274–284, 2005; Stadler, R.H., Acrylamide formation in different foods and potential strategies for reduction, Adv. Exp. Med. Biol. 561, 157–169, 2005; Lopachin, R.M. and Decaprio, A.P., Protein adduct formation as a molecular mechanism in neurotoxicity, Toxicol. Sci. 86, 214–225, 2005.


Mandel, P. and DeFeudis, F.V., Eds., GABA—Biochemistry and CNS Functions, Plenum Press, New York, 1979; Costa, E. and Di Chiara, G., GABA and Benzodiazepine Receptors, Raven Press, New York, 1981; Racagni, G. and Donoso, A.O., GABA and Endocrine Function, Raven Press, New York, 1986; Squires, R.F., GABA and Benzodiazepine Receptors, CRC Press, Boca Raton, FL, 1988; Martin, D.L. and Olsen, R.W., GABA in the Nervous System: The View at Fifty Years, Lippincott, Williams & Wilkins, Philadelphia, PA, 2000.


Benos, D.J., A molecular probe of sodium transport in tissues and cells, Am. J. Physiol. 242, C131–C145, 1982; Garty, H., Molecular properties of epithelial, amiloride-blockable Na+ channels, FASEB J. 8, 522–528, 1994; Barbry, P. and Lazdunski, M., Structure and regulation of the amiloride-sensitive epithelial sodium channel, Ion Channels 4, 115–167, 1996; Kleyman, T.R., Sheng, S., Kosari, F., and Kieber-Emmons, T., Mechanism of action of amiloride: a molecular perspective, Semin. Nephrol. 19, 524–532, 1999; Alvarez de la Rosa, D., Canessa, C.M., Fyfe, G.K., and Zhang, P., Structure and regulation of amiloride-sensitive sodium channels, Annu. Rev. Physiol. 62, 573–594, 2000; Haddad, J.J., Amiloride and the regulation of NF-κβ: an unsung crosstalk and missing link between fluid dynamics and oxidative stress-related inflammation — controversy or pseudo-controversy, Biochem. Biophys. Res. Commun. 327, 373–381, 2005.


Hase, S., Hara, S., and Matsushima, Y., Tagging of sugars with a fluorescent compound, 2-aminopyridine, J. Biochem. 85, 217–220, 1979; Hase, S., Ibuki, T., and Ikenaka, T., Reexamination of the pyridylamination used for fluorescence labeling of oligosaccharides and its application to glycoproteins, J. Biochem. 95, 197–203, 1984; Chen, C. and Zheng, X., Development of the new antimalarial drug pyronaridine: a review, Biomed. Environ. Sci. 5, 149–160, 1992; Hase, S., Analysis of sugar chains by pyridylamination, Methods Mol. Biol. 14, 69–80, 1993; Oefner, P.J. and Chiesa, C., Capillary electrophoresis of carbohydrates, Glycobiology 4, 397–412, 1994; Dyukova, V.I., Shilova, N.V., Galanina, O.E. et al., Design of carbohydrate multiarrays, Biochim. Biophys. Acta 1760, 603–609, 2006; Takegawa, Y., Deguchi, K., Keira, T. et al., Separation of isomeric 2-aminopyridine derivatized N-glycans and N-glycopeptides of human serum immunoglobulin G by using a zwitterionic type of hydrophilic-interaction chromatography, J. Chromatog. A 1113, 177–181, 2006; Suzuki, S., Fujimori, T., and Yodoshi, M., Recovery of free oligosaccharides from derivatives labeled by reductive amination, Anal. Biochem. 354, 94–103, 2006; Caballero, N.A., Melendez, F.J., Munoz-Caro, C., and Nino, A., Theoretical prediction of relative and absolute pK(a) values of aminopyridine, Biophys. Chem., 124, 155–160, 2006.


Gibbons, G.R., Page, J.D., and Chaney, S.G., Treatment of DNA with ammonium bicarbonate or thiourea can lead to underestimation of platinum-DNA monoadducts, Cancer Chemother. Pharmacol. 29, 112–116, 1991; Sorenson, S.B., Sorenson, T.L., and Breddam, K., Fragmentation of protein by S. aureus strain V8 protease. Ammonium bicarbonate strongly inhibits the enzyme but does not improve the selectivity for glutamic acid, FEBS Lett. 294, 195–197, 1991; Fichtinger-Schepman, A.M., van Dijk-Knijnenburg, H.C., Dijt, F.J. et al., Effects of thiourea and ammonium bicarbonate on the formation and stability of bifunctional cisplatinin-DNA adducts: consequences for the accurate quantification of adducts in (cellular) DNA, J. Inorg. Biochem. 58, 177–191, 1995; Overcashier, D.E., Brooks, D.A., Costantino, H.R., and Hus, C.C., Preparation of excipient-free recombinant human tissue-type plasminogen activator by lyophilization from ammonium bicarbonate solution: an investigation of the two-stage sublimation process, J. Pharm. Sci. 86, 455–459, 1997.


Ferguson, R.N., Edelhoch, H., Saroff, H.A. et al., Negative cooperativity in the binding of thyroxine to human serum prealbumin. Preparation of tritium-labeled 8-anilino-1-naphthalenesulfonic acid, Biochemistry 14, 282–289, 1975; Ogasahara, K., Koike, K., Hamada, M., and Hiraoka, T., Interaction of hydrophobic probes with the apoenzyme of pig heart lipoamide dehydrogenase, J. Biochem. 79, 967–975, 1976; De Campos Vidal, B., The use of the fluorescence probe 8-anilinonaphthalene sulfate (ANS) for collagen and elastin histochemistry, J. Histochem. Cytochem. 26, 196–201, 1978; Royer, C.A., Fluorescence spectroscopy, Methods Mol. Biol. 40, 65–89, 1995; Celej, M.S., Dassie, S.A., Freire, E. et al., Ligand-induced thermostability in proteins: thermodynamic analysis of ANS-albumin interaction, Biochim. Biophys. Acta 1750, 122–133, 2005; Banerjee, T. and Kishore, N., Binding of 8-anilinonaphthalene sulfonate to dimeric and tetrameric concanavalin A: energetics and its implications on saccharide binding studied by isothermal titration calorimetry and spectroscopy, J. Phys. Chem. B Condens. Matter Mater. Surf. Interfaces Biophys. 110, 7022–7028, 2006; Sahu, K., Mondal, S.K., Ghosh, S. et al., Temperature dependence of salvation dynamics and anisotropy decay in a protein: ANS in bovine serum albumin, J. Chem. Phys. 124, 124909, 2006; Wang, G., Gao, Y., and Geng, M.L., Analysis of heterogeneous fluorescence decays in proteins. Using fluorescence lifetime of 8-anilino-1-naphthalenesulfonate to probe apomyoglobin unfolding at equilibrium, Biochim. Biophys. Acta 1760, 1125–1137, 2006; Greene, L.H., Wijesinha-Bettoni, R., and Redfield, C., Characterization of the molten globule of human serum retinol-binding protein using NMR spectroscopy, Biochemistry 45, 9475–9484, 2006.


Moncada, S. and Vane, J.R., Interaction between anti-inflammatory drugs and inflammatory mediators. A reference to products of arachidonic acid metabolism, Agents Actions Suppl. 3, 141–149, 1977; Moncada, S. and Higgs, E.A., Metabolism of arachidonic acid, Ann. N.Y. Acad. Sci. 522, 454–463, 1988; Piomelli, D., Arachidonic acid in cell signaling, Curr. Opin. Cell Biol. 5, 274–280, 1993; Janssen-Timmen, U., Tomic, I., Specht, E. et al., The arachidonic acid cascade, eicosanoids, and signal transduction, Ann. N.Y. Acad. Sci. 733, 325–334, 1994; Wang, X. and Stocco, D.M., Cyclic AMP and arachidonic acid: a tale of two pathways, Mol. Cell. Endocrinol. 158, 7–12, 1999; Brash, A.R., Arachidonic acid as a bioactive molecule, J. Clin. Invest. 107, 1339–1345, 2001; Luo, M., Flamand, N., and Brock, T.G., Metabolism of arachidonic acid to eicosanoids within the nucleus, Biochim. Biophys. Acta 1761, 618–625, 2006; Balboa, M.A. and Balsinde, J., Oxidative stress and arachidonic acid mobilization, Biochim. Biophys. Acta 1761, 385–391, 2006.


Barnes, M.J. and Kodicek, E., Biological hydroxylations and ascorbic acid with special regard to collagen metabolism, Vitam. Horm. 30, 1–43, 1972; Leibovitz, B. and Siegel, B.V., Ascorbic acid and the immune response, Adv. Exp. Med. Biol. 135, 1–25, 1981; Englard, S. and Seifter, S., The biochemical functions of ascorbic acid, Annu. Rev. Nutr. 6, 365–406, 1986; Levine, M. and Hartzell, W., Ascorbic acid: the concept of optimum requirements, Ann. N.Y. Acad. Sci. 498, 424–444, 1987; Padh, H., Cellular functions of ascorbic acid, Biochem. Cell Biol. 68, 1166–1173, 1990; Meister, A., On the antioxidant effects of ascorbic acid and glutathione, Biochem. Pharmacol. 44, 1905–1915, 1992; Wolf, G., Uptake of ascorbic acid by human neutrophils, Nutr. Rev. 51, 337–338, 1993; Kimoto, E., Terada, S., and Yamaguchi, T., Analysis of ascorbic acid, dehydroascorbic acid, and transformation products by ion-pairing high-performance liquid chromatography with multiwavelength ultraviolet and electrochemical detection, Methods Enzymol. 279, 3–12, 1997; May, J.M., How does ascorbic acid prevent endothelial dysfunction? Free Rad. Biol. Med. 28, 1421–1429, 2000; Smirnoff, N. and Wheeler, G.L., Ascorbic acid in plants: biosynthesis and function, Crit. Rev. Biochem. Mol. Biol. 35, 291–314, 2000; Arrigoni, O. and De Tullio, M.C., Ascorbic acid: much more than just an antioxidant, Biochim. Biophys. Acta 1569, 1–9, 2002; Akyon, Y., Effect of antioxidant on the immune response of Helicobacter pyrlori, Clin. Microbiol. Infect. 8, 438–441, 2002; Takanaga, H., MacKenzie, B., and Hediger, M.A., Sodium-dependent ascorbic acid transporter family SLC23, Pflügers Arch. 447, 677–682, 2004.


Chalmers, R.M., Keen, J.N., and Fewson, C.A., Comparison of benzyl alcohol dehydrogenases and benzaldehyde dehydrogenases from the benzyl alcohol and mandelate pathways in Acinetobacter calcoaceticus and the TOL-plasmid-encoded toluene pathway in Pseudomonas putida. N-terminal amino acid sequences, amino acid composition, and immunological cross-reactions, Biochem. J. 273, 99–107, 1991; Pettersen, E.O., Larsen, R.O., Borretzen, B. et al., Increased effect of benzaldehyde by exchanging the hydrogen in the formyl group with deuterium, Anticancer Res. 11, 369–373, 1991; Nierop Groot, M.N. and de Bont, J.A.M., Conversion of phenylalanine to benzaldehyde initiated by an amino-transferase in Lactobacillus plantarum, Appl. Environ. Microbiol. 64, 3009–3013, 1998; Podyminogin, M.A., Lukhtanov, E.A., and Reed, M.W., Attachment of benzaldehyde-modified oligodeoxynucleotide probes to semicarbazide-coated glass, Nucleic Acids Res. 29, 5090–5098, 2001; Kurchan, A.N. and Kutateladze, A.G., Amino acid-based dithiazines: synthesis and photofragmentation of their benzaldehyde adducts, Org. Lett. 4, 4129–4131, 2002; Kneen, M.M., Pogozheva, I.D., Kenyon, G.L., and McLeish, M.J., Exploring the active site of benzaldehyde lyase by modeling and mutagenesis, Biochim. Biophys. Acta 1753, 263–271, 2005; Mosbacher, T.G., Mueller, M., and Schultz, G.E., Structure and mechanism of the ThDP-dependent benzaldehyde lyase from Pseudomonas fluorescens, FEBS J. 272, 6067–6076, 2005; Sudareva, N.N. and Chubarova, E.V., Time-dependent conversion of benzyl alcohol to benzaldehyde and benzoic acid in aqueous solution, J. Pharm. Biomed. Anal. 41, 1380–1385, 2006.


Ensinck, J.W., Shepard, C., Dudl, R.J., and Williams, R.H., Use of benzamidine as a proteolytic inhibitor in the radioimmunoassay of glucagon in plasma, J. Clin. Endocrinol. Metab. 35, 463–467, 1972; Bode, W. and Schwager, P., The refined crystal structure of bovine beta-trypsin at 1.8 Å resolution. II. Crystallographic refinement, calcium-binding site, benzamidine-binding site and active site at pH 7.0., J. Mol. Biol. 98, 693–717, 1975; Nastruzzi, C., Feriotto, G., Barbieri, R. et al., Differential effects of benzamidine derivatives on the expression of c-myc and HLA-DR alpha genes in a human B-lymphoid tumor cell line, Cancer Lett. 38, 297–305, 1988; Clement, B., Schmitt, S., and Zimmerman, M., Enzymatic reduction of benzamidoxime to benzamidine, Arch. Pharm. 321, 955–956, 1988; Clement, B., Immel, M., Schmitt, S., and Steinman, U., Biotransformation of benzamidine and benzamidoxime in vivo, Arch. Pharm. 326, 807–812, 1993; Renatus, M., Bode, W., Huber, R. et al., Structural and functional analysis of benzamidine-based inhibitors in complex with trypsin: implications for the inhibition of factor Xa, tPA, and urokinase, J. Med. Chem. 41, 5445–5456, 1998; Henriques, R.S., Fonseca, N., and Ramos, M.J., On the modeling of snake venom serine proteinase interactions with benzamidine-based thrombin inhibitors, Protein Sci. 13, 2355–2369, 2004; Gustavsson, J., Farenmark, J., and Johansson, B.L., Quantitative determination of the ligand content in benzamidine Sepharose® 4 Fast Flow media with ion-pair chromatography, J. Chromatog. A 1070, 103–109, 2005.


Lovley, D.R., Anaerobic benzene degradation, Biodegradation 11, 107–116, 2000; Snyder, R., Xenobiotic metabolism and the mechanism(s) of benzene toxicity, Drug Metab. Rev. 36, 531–547, 2004; Rana, S.V. and Verma, Y., Biochemical toxicity of benzene, J. Environ. Biol. 26, 157–168, 2005; Lin, Y.S., McKelvey, W., Waidyanatha, S., and Rappaport, S.M., Variability of albumin adducts of 1,4-benzoquinone, a toxic metabolite of benzene, in human volunteers, Biomarkers 11, 14–27, 2006; Baron, M. and Kowalewski, V.J., The liquid water-benzene system, J. Phys. Chem. A Mol. Spectrosc. Kinet. Environ. Gen. Theory 100, 7122–7129, 2006; Chambers, D.M., McElprang, D.O., Waterhouse, M.G., and Blount, B.C., An improved approach for accurate quantiation of benzene, toluene, ethylbenzene, zylene, and styrene in blood, Anal. Chem. 78, 5375–5383, 2006.


Ahlquist, D.A. and Schwartz, S., Use of leuco-dyes in the quantitative colorimetric microdetermination of hemoglobin and other heme compounds, Clin. Chem. 21, 362–369, 1975; Josephy, P.D., Benzidine: mechanisms of oxidative activation and mutagensis, Fed. Proc. 45, 2465–2470, 1986; Choudhary, G., Human health perspectives on environmental exposure to benzidine: a review, Chemosphere 32, 267–291, 1996; Madeira, P., Nunes, M.R., Borges, C. et al., Benzidine photodegradation: a mass spectrometry and UV spectroscopy combined study, Rapid Commun. Mass Spectrom. 19, 2015–2020, 2005; Saitoh, T., Yoshida, S., and Ichikawa, J., Naphthalene-1,8-diylbis(diphenylmethylium) as an organic two-electron oxidant: benzidine synthesis via oxidative self-coupling of N,N-dialkylanilines, J. Org. Chem. 71, 6414–6419, 2006.


Bonelli, F.S. and Jonas, A., Reaction of lecithin: cholesterol acyltransferase with a water-soluble substrate: effects of surfactants, Biochim. Biophys. Acta 1166, 92–98, 1993; Aigner, A., Jager, M., Pasternack, R. et al., Purification and characterization of cysteine-S-conjugate N-acetyltransferase from pig kidney, Biochem. J. 317, 213–218, 1996; Mechref, Y. and Eirassi, Z., Micellar electrokinetic capillary chromatography with in-situ charged micelles. 4. Evaluation of novel chiral micelles consisting of steroidal glycoside surfactant borate complexes, J. Chromatog. A 724, 285–296, 1996; Abe, S., Kunii, S., Fujita, T., and Hiraiwa, K., Detection of human seminal gamma-glutamyl transpeptidase in stains using sandwich ELISA, Forensic Sci. Int. 91, 19–28, 1998; Akutsu, Y., Nakajima-Kambe, T., Nomura, N., and Nakahara, T., Purification and properties of a polyester polyurethane-degrading enzyme form Comamonas acidovorans TB-35, Appl. Environ. Microbiol. 64, 62–67, 1998: Connor, R.J., Engler, H., Machemer, T. et al., Identification of polyamides that enhance adenovirus-mediated gene expression in the urothelium, Gene Therapy 8, 41–48, 2001; Vajdos, F.F., Ultsch, M., Schaffer, M.L. et al., Crystal structure of human insulin-like growth factor-1: detergent binding inhibits binding protein interactions, Biochemistry 40, 11022–11029, 2001; Kuball, J., Wen, S.F., Leissner, J. et al., Successful adenovirus-mediated wild-type p53 gene transfer in patients with bladder cancer by intravesical vector instillation, J. Clin. Oncol. 20, 957–965, 2002; Susasara, K.M., Xia, F., Gronke, R.S., and Cramer, S.M., Application of hydrophobic interaction displacement chromatography for an industrial protein purification, Biotechnol. Bioeng. 82, 330–339, 2003; Ishibashi, A. and Nakashima, N., Individual dissolution of single-walled carbon nanotubes in aqueous solutions of steroid or sugar compounds and their Raman and near-IR spectral properties, Chemistry, 12, 7595–7602, 2006.


Knappe, J., Mechanism of biotin action, Annu. Rev. Biochem. 39, 757–776, 1970; Dunn, M.J., Detection of proteins on blots using the avidin-biotin system, Methods Mol. Biol. 32, 227–232, 1994; Wisdom, G.B., Enzyme and biotin labeling of antibody, Methods Mol. Biol. 32, 433–440, 1994; Wilbur, D.S., Pathare, P.M, Hamlin, D.K. et al., Development of new biotin/streptavidin reagents for pretargeting, Biomol. Eng. 16, 113–118, 1999; Jitrapakdee, S. and Wallace, J.C., The biotin enzyme family: conserved structural motifs and domain rearrangements, Curr. Protein Pept. Sci. 4, 217–229, 2003; Nikolau, B.J., Ohlrogge, J.B., and Wurtels, E.S., Plant biotin-containing carboxylases, Arch. Biochem. Biophys. 414, 211–222, 2003; Fernandez-Mejia, C., Pharmacological effects of biotin, J. Nutri. Biochem. 16, 424–427, 2005; Wilchek, M., Bayer, E.A., and Livnah, O., Essentials of biorecognition: the (strept)avidin-biotin system as a model for protein–protein and protein–ligand interactions, Immunol. Lett. 103, 27–32, 2006; Furuyama, T. and Henikoff, S., Biotin-tag affinity purification of a centromeric nucleosome assembly complex, Cell Cycle 5, 1269–1274, 2006; Streaker, E.D. and Beckett, D., Nonenzymatic biotinylation of a biotin carboxyl carrier protein: unusual reactivity of the physiological target lysine, Protein Sci. 15, 1928–1935, 2006; Raichur, A.M., Voros, J., Textor, M., and Fery, A., Adhesion of polyelectrolyte microcapsules through biotin-streptavidin specific interaction, Biomacromolecules 7, 2331–2336, 2006. For biotin switch assay, see Martinez-Ruiz, A. and Lamas, S., Detection and identification of S-nitrosylated proteins in endothelial cells, Methods Enzymol. 396, 131–139, 2005; Huang, B. and Chen, C., An ascorbate-dependent artifact that interferes with the interpretation of the biotin switch assay, Free Radic. Biol. Med. 41, 562–567, 2006; Gladwin, M.T., Wang, X., and Hogg, N., Methodological vexation about thiol oxidation versus S-nitrosation — a commentary on “An ascorbate-dependent artifact that interferes with the interpretation of the biotin-switch assay,” Free Radic. Biol. Med. 41, 557–561, 2006.


Jensen, H.L. and Schroder, M., Urea and biuret as nitrogen sources for Rhizobium spp., J. Appl. Bacteriol. 28, 473–478, 1965; Ronca, G., Competitive inhibition of adenosine deaminase by urea, guanidine, biuret, and guanylurea, Biochim. Biophys. Acta 132, 214–216, 1967; Oltjen, R.R., Slyter, L.L., Kozak, A.S., and Williams, E.E., Jr., Evaluation of urea, biuret, urea phosphate, and uric acid as NPN sources for cattle, J. Nutr. 94, 193–202, 1968; Tsai, H.Y. and Weber, S.G., Electrochemical detection of oligopeptides through the precolumn formation of biuret complexes, J. Chromatog. 542, 345–350, 1991; Gawron, A.J. and Lunte, S.M., Optimization of the conditions for biuret complex formation for the determination of peptides by capillary electrophoresis with ultraviolet detection, Clin. Chem. 51, 1411–1419, 2000; Roth, J., O’Leary, D.J., Wade, C.G. et al., Conformational analysis of alkylated biuret and triuret: evidence for helicity and helical inversion in oligoisocyates, Org. Lett. 2, 3063–3066, 2000; Hortin, G.L., and Mellinger, B., Cross-reactivity of amino acids and other compounds in the biuret reaction: interference with urinary peptide measurements, Clin. Chem. 51, 1411–1419, 2005.



Litteria, M. and Recknagel, R.O., A simplified blue tetrazolium reaction, J. Lab. Clin. Med. 48, 463–468, 1955; Sinsheimer, J.E. and Salim, E.F., Reactivity of blue tetrazolium with nonketol compounds, Anal. Chem. 37, 566–569, 1965; Graham, R.E., Biehl, E.R., Kenner, C.T. et al., Reduction of blue tetrazolium by corticosteroids, J. Pharm. Sci. 64, 226–230, 1975; Baba, N., Burtubise, P., and Myser, T., Immunofluorescence and immunoperoxidase observations of anti-lactic dehydrogenase-1 antibody, J. Histochem. Cytochem. 24, 572–577, 1976; Biehl, E., Wooten, R., Kenner, C.T., and Graham, R.E., Kinetic and mechanistic studies of blue tetrazolium reaction with phenylhydrazines, J. Pharm. Sci. 67, 927–930, 1978; Van Noorden, C.J., Tas, J., and Vogels, I.M., Cytophotometry of glucose-6-phosphate dehydrogenase activity in individual cells, Histochem. J. 15, 583–599, 1983; Maravelias, C., Dona, A., Athanaselis, S., and Koutselinis, A., The importance of performing in vitro cytotoxicity testing before immodulation evaluation, Vet. Hum. Toxicol. 42, 292–296, 2000; Reddy, R.M., Tsai, W.S., Ziauddin, M.F. et al., Cisplatin enhances apoptosis induced by a tumor-selective adenovirus-expressing tumor necrosis factor-related apoptosis-inducing ligand, J. Thorac. Cardiovasc. Surg. 128, 883–891, 2004.


Sciarra, J.J. and Monte Bovi, A.J., Study of the boric acid–glycerin complex. II. Formation of the complex at elevated temperature, J. Pharm. Sci. 51, 238–242, 1962; Walborg, E.F., Jr. and Lantz, R.S., Separation and quantitation of saccharides by ion-exchange chromatography utilizing boric acid–glycerol buffers, Anal. Biochem. 22, 123–133, 1968; Lerch, B. and Stegemann, H., Gel electrophoresis of proteins in borate buffer. Influence of some compounds complexing with boric acid, Anal. Biochem. 29, 76–83, 1969; Walborg, E.F., Jr., Ray, D.B., and Ohrberg, L.E., Ion-exchange chromatography of saccharides: an improved system utilizing boric acid/2,3-butanediol buffers, Anal. Biochem. 29, 433–440, 1969; Chen, F.T. and Sternberg, J.C., Characterization of proteins by capillary electrophoresis in fused-silica columns: review on serum protein anlaysis and application to immunoassays, Electrophoresis 15, 13–21, 1994; Allen, R.C. and Doktycz, M.J., Discontinuous electrophoresis revisited: a review of the process, Appl. Theor. Electrophor. 6, 1–9, 1996; Manoravi, P., Joseph, M., Sivakumar, N., and Balasubramanian, H., Determination of isotopic ratio of boron in boric acid using laser mass spectrometry, Anal. Sci. 21, 1453–1455, 2005; De Muynck, C., Beauprez, J., Soetaert, W., and Vandamme, E.J., Boric acid as a mobile phase additive for high-performance liquid chromatography separation of ribose, arabinose, and ribulose, J. Chromatog. A 1101, 115–121, 2006; Herrmannova, M., Kirvankova, L., Bartos, M., and Vytras, K., Direct simultaneous determination of eight sweeteners in foods by capillary isotachophoresis, J. Sep. Sci. 29, 1132–1137, 2006; Alencar de Queiroz, A.A., Abraham, G.A., Pires Camillo, M.A. et al., Physicochemical and antimicrobial properties of boron-complexed polyglycerol-chitosan dendrimers, J. Biomater. Sci. Polym. Ed. 17, 689–707, 2006; Ringdahl, E.N., Recurrent vulvovaginal candidiasis, Mol. Med. 103, 165–168, 2006.


Boulanger, P., Lemay, P., Blair, G.E., and Russell, W.C., Characteriztion of adenovirus protein IX, J. Gen. Virol. 44, 783–800, 1979; Russell, J., Kathendler, J., Kowalski, K. et al., The single tryptophan residue of human placental lactogen. Effects of modification and cleavage on biological activity and protein conformation, J. Biol. Chem. 256, 304–307, 1981; Moskaitis, J.E. and Campagnoni, A.T., A comparison of the dodecyl sulfate-induced precipitation of the myelin basic protein with other water-soluble proteins, Neurochem. Res. 11, 299–315, 1986; Mahboub, S., Richard, C., Delacourte, A., and Han, K.K., Applications of chemical cleavage procedures to the peptide mapping of neurofilament triplet protein bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Anal. Biochem. 154, 171–182, 1986; Rahali, V. and Gueguen, J., Chemical cleavage of bovine beta-lactoglobulin by BPNS-skatole for preparative purposes: comparative study of hydrolytic procedure and peptide characterization, J. Protein Chem. 18, 1–12, 1999; Swamy, N., Addo, J., Vskokovic, M.R., and Ray, R., Probing the vitamin D sterol-binding pocket of human vitamin D-binding protein with bromoacetate affinity-labeling reagents containing the affinity probe at C-3, C-6, C-11, and C-19 positions of parent vitamin D sterols, Arch. Biochem. Biophys. 373, 471–478, 2000; Celestina, F. and Suryanarayana, T., Biochemical characterization and helix-stabilizing properties of HSNP-C’ from the thermophilic archaeon Sulfolobus acidocaldarius, Biochem. Biophys. Res. Commun. 267, 614–618, 2000; Kibbey, M.M., Jameson, M.J., Eaton, E.M., and Rosenzweig, S.A., Insulinlike growth factor binding protein-2: contributions of the C-terminal domain to insulinlike growth factor-1 binding, Mol. Pharmacol. 69, 833–845, 2006.


Glick, D.M., Goren, H.J., and Barnard, E.A., Concurrent bromoacetate reaction at histidine and methionine residues in ribonuclease, Biochem. J. 102, 7C–10C, 1967; Goren, H.J. and Barnard, E.A., Relation of reactivity to structure in pancreatic ribonuclease. I. An analysis of the various reactions with bromoacetate in the pH range of 2–7, Biochemistry 9, 959–973, 1970; Goren, H.J. and Barnard, E.A., Relation of reactivity to structure in pancreatic ribonuclease. II. Positions of residues alkylated in certain conditions by bromoacetate, Biochemistry 9, 974–983, 1970; Lennette, E.P. and Plapp, B.V., Kinetics of carboxymethylation of histidine hydantoin, Biochemistry 18, 3933–3938, 1979; Adamczyk, M., Gebler, J.C., and Wu, J., A simple method to identify cysteine residues by isotopic labeling and ion trap mass spectrometry, Rapid Commun. Mass Spectrom. 13, 1813–1817, 1999; Schelte, P., Boeckler, C., Frisch, B., and Schuber, F., Differential reactivity of maleimide and bromoacetyl functions with thiols: application to the preparation of lysosomal diepitope constructs, Bioconjug. Chem. 11, 118–123, 2000; Filmon, R., Grizon, F., Basle, M.F., and Chappaard, D., Effects of negatively charged groups (carboxymethyl) on the calcification of poly(2-hydroxyethyl methacrylate), Biomaterials 23, 3053–3059, 2002; Barron, L. and Paull, B., Direct detection of trace haloacetates in drinking water using microbore ion chromatography. Improved detector sensitivity using a hydroxide gradient and a monolithic ion-exchange type suppressor, J. Chromatog. A 1047, 205–212, 2004; Zhang, L., Arnold, W.A., and Hozalski, R.M., Kinetics of haloacetic acid reactions with Fe(0), Environ. Sci. Technol. 38, 6881–6889, 2004; Lee, S. and Perez-Luna, V.H., Dextran-gold nanoparticle hybrid material for biomolecule immobilization and detection, Anal. Chem. 77, 7204–7211, 2005.


Erlanger, B.F., Vratrsanos, S.M., Wasserman, N., and Cooper, A.G., A chemical investigation of the active center of pepsin, Biochem. Biophys. Res. Commun. 23, 243–245, 1966; Yang, C.C. and King, K., Chemical modification of the histidine residue in basic phospholipase A2 from the venom of Naja nigricollis, Biochim. Biophys. Acta. 614, 373–388, 1980; Darke, P.L., Jarvis, A.A., Deems, R.A., and Dennis, E.A., Further characterization and N-terminal sequence of cobra venom phospholipase A2, Biochim. Biophys. Acta 626, 154–161, 1980; Ackerman, S.K., Matter, L., and Douglas, S.D., Effects of acid proteinase inhibitors on human neutrophil chemotaxis and lysosomal enzyme release. II. Bromophenacyl bromide and 1,2-epoxy-3-(p-nitrophenoxy)propane, Clin. Immunol. Immunopathol. 26, 213–222, 1983; Carine, K. and Hudig, D., Assessment of a role for phospholipase A2 and arachidonic acid metabolism in human lymphocyte natural cytotoxicity, Cell Immunol. 87, 270–283, 1984; Duque, R.E., Fantone, J.C., Kramer, C. et al., Inhibition of neutrophil activation by p-bromophenacyl bromide and its effects on phospholipase A2, Br. J. Pharmacol. 88, 463–472, 1986; Zhukova, A., Gogvadze, G., and Gogvadze, V., p-bromophenacyl bromide prevents cumene hydroperoxide-induced mitochondrial permeability transition by inhibiting pyridine nucleotide oxidation, Redox Rep. 9, 117–121, 2004; Thommesen, L. and Laegreid, A., Distinct differences between TNF receptor 1- and TNR receptor 2-mediated activation of NF-κβ, J. Biochem. Mol. Biol. 38, 281–289, 2005; Yue, H.Y., Fujita, T., and Kumamoto, E., Phospholipase A2 activation by melittin enhances spontaneous glutamatergic excitatory transmission in rat substantia gelatinosa neurons, Neuroscience 135, 485–495, 2005; Costa-Junior, H.M., Hamaty, F.C., de Silva Farias, R. et al., Apoptosis-inducing factor of a cytotoxic T-cell line: involvement of a secretory phospholipase A(2), Cell Tissue Res. 324, 255–266, 2006; Marchi-Salvador, D.P., Fernandes, C.A., Amui, S.F. et al., Crystallization and preliminary X-ray diffraction analysis of a myotoxic Lys49-PLA2 from Bothrops jararacussu venom complexed with p-bromophenacyl bromide, Acta Crystallograph. Sect. F Struct. Biol. Cryst. Commun. 62, 600–603, 2006.


Schilling, K. and Waldmann-Meyer, H., The interaction of bromophenol blue with serum albumin and gamma-globulin in acid medium, Arch. Biochem. Biophys. 64, 291–301, 1956; Cohen, A.H., Temperature jump studies of the binding of bromophenol blue to beta-lactoglobulin in the vicinity of the N–R transition, J. Biol. Chem. 245, 738–745, 1970; Harruff, R.C. and Jenkins, W.T., The binding of bromophenol blue to aspartate aminotransferase, Arch. Biochem. Biophys. 176, 206–213, 1976; Mitchell, J.P., Model system studies of staining procedures for lysine and arginine residues, Histochemistry 52, 151–157, 1977; Asao, T., Quantitative analysis of proteins by the use of SDS-polyacrylamide-gel electrophoresis, Anal. Biochem. 77, 321–331, 1977; Greenberg, C.S. and Craddock, P.R., Rapid single-step membrane protein assay, Clin. Chem. 28, 1725–1726, 1982; Bertsch, M. and Kassner, R.J., Selective staining of proteins with hydrophobic surface sites on a native electrophoretic gel, J. Proteome Res. 2, 469–475, 2003; Li, J., Chatterjee, K., Medek, A. et al., Acid-base characteristics of bromophenol blue-citrate buffer systems in the amorphous state, J. Pharm. Sci. 93, 697–712, 2004; Haritoglou, C., Yu, A., Freyer, W. et al., An evaluation of novel dyes for intraocular surgery, Invest. Ophthalmol. Vis. Sci. 46, 3315–3322, 2005; Haritoglou, C., Tadayoni, R., May, C.A. et al., Short-term in vivo evaluation of novel vital dyes for interocular surgery, Retina 26, 673–678, 2006; Schuettauf, F., Haritoglou, C., and May, C.A., Administration of novel dyes for intraocular surgery: an in vivo toxicity animal study, Invest. Ophthalmol. Vis. Sci. 47, 3573–3578, 2006; Zeroual, Y., Kim, B.S., Kim, C.S. et al., A comparative study on biosorption characteristics of certain fungi for bromophenol blue dye, Appl. Biochem. Biotechnol. 134, 51–60, 2006.


McAlpine, J.C., Histochemical demonstration of the activation of rat acetylcholinesterase by sodium cacodylate and cacodylic acid using the thioacetic acid method, J. R. Microsc. Soc. 82, 95–106, 1963; Jacobson, K.B., Murphy, J.B., and Das Sarma, B., Reaction of cacodylic acid with organic thiols, FEBS Lett. 22, 80–82, 1972; Travers, F., Douzou, P., Pederson, T., and Gunsalus, I.C., Ternary solvents to investigate proteins at subzero temperature, Biochimie 57, 43–48, 1975; Young, C.W., Dessources, C., Hodas, S., and Bittar, E.S., Use of cationic disc electrophoresis near neutral pH in the evaluation of trace proteins in human plasma, Cancer Res. 35, 1991–1995, 1975; Chirpich, T.P., The effect of different buffers on terminal deoxynucleotidyl transferase activity, Biochim. Biophys. Acta 518, 535–538, 1978; Nunes, J.F., Aguas, A.P., and Soares, J.O., Growth of fungi in cacodylate buffer, Stain Technol. 55, 191–192, 1980; Caswell, A.H. and Bruschwig, J.P., Identification and extraction of proteins that compose the triad junction of skeletal muscle, J. Cell Biol. 99, 929–939, 1984; Parks, J.C. and Cohen, G.M., Glutaraldehyde fixatives for preserving the chick’s inner ear, Acta Otolaryngol. 98, 72–80, 1984; Song, A.H. and Asher, S.A., Internal intensity standards for heme protein UV resonance Raman studies: excitation profiles of cacodylic acid and sodium selenate, Biochemistry 30, 1199–1205, 1991; Henney, P.J., Johnson, E.L., and Cothran, E.G., A new buffer system for acid PAGE typing of equine protease inhibitor, Anim. Genet. 25, 363–364, 1994; Jezewska, M.J., Rajendran, S., and Bujalowski, W., Interactions of the 8-kDa domain of rat DNA polymerase beta with DNA, Biochemistry 40, 3295–3307, 2001; Kenyon, E.M. and Hughes, M.F., A concise review of the toxicity and carcinogenicity of dimethylarsinic acid, Toxicology 160, 227–236, 2001; Cohen, S.M., Arnold, L.L., Eldan, M. et al., Methylated arsenicals: the implications of metabolism and carcinogenicity studies in rodents to human risk management, Crit. Rev. Toxicol. 99–133, 2006.

Calcium Chloride

CaCl2; Various Hydrates


Anhydrous form as drying agent for organic solvents, variety of manufacturing uses; meat quality enhancement; therapeutic use in electrolyte replacement and bone cements; source of calcium ions for biological assays.

Barratt, J.O., Thrombin and calcium chloride in relation to coagulation, Biochem. J. 9, 511–543, 1915; Van der Meer, C., Effect of calcium chloride on choline esterase, Nature 171, 78–79, 1952; Bhat, R. and Ahluwalia, J.C., Effect of calcium chloride on the conformation of proteins. Thermodynamic studies of some model compounds, Int. J. Pept. Protein Res. 30, 145–152, 1987; Furihata, C., Sudo, K., and Matsushima, T., Calcium chloride inhibits stimulation of replicative DNA synthesis by sodium chloride in the pyloric mucosa of rat stomach, Carcinogenesis 10, 2135–2137, 1989; Ishikawa, K., Ueyama, Y., Mano, T. et al., Self-setting barrier membrane for guided tissue regeneration method: initial evaluation of alginate membrane made with sodium alginate and calcium chloride aqueous solutions, J. Biomed. Mater. Res. 47, 111–115, 1999; Vujevic, M., Vidakovic-Cifrek, Z., Tkalec, M. et al., Calcium chloride and calcium bromide aqueous solutions of technical and analytical grade in Lemna bioassay, Chemosphere 41, 1535–1542, 2000; Miyazaki, T., Ohtsuki, C., Kyomoto, M. et al., Bioactive PMMA bone cement prepared by modification with methacryloxypropyltrimethoxysilane and calcium chloride, J. Biomed. Mater. Res. A 67, 1417–1423, 2003; Harris, S.E., Huff-Lonegan, E., Lonergan, S.M. et al., Antioxidant status affects color stability and tenderness of calcium chloride-injected beef, J. Anim. Sci. 79, 666–677, 2001; Behrends, J.M., Goodson, K.J., Koohmaraie, M. et al., Beef customer satisfaction: factors affecting consumer evaluations of calcium chloride-injected top sirloin steaks when given instructions for preparation, J. Anim. Sci. 83, 2869–2875, 2005.


Laurent, T.C. and Scott, J.E., Molecular weight fractionation of polyanions by cetylpyridinium chloride in salt solutions, Nature 202, 661–662, 1964; Kiss, A., Linss, W., and Geyer, G., CPC-PTA section staining of acid glycans, Acta Histochem. 64, 183–186, 1979; Khan, M.Y. and Newman, S.A., An assay for heparin by decrease in color yield (DECOY) of a protein-dye-binding reaction, Anal. Biochem. 187, 124–128, 1990; Chardin, H., Septier, D., and Goldberg, M., Visualization of glycosaminoglycans in rat incisor predentin and dentin with cetylpyridinium chloride-glutaraldehyde as fixative, J. Histochem. Cytochem. 38, 885–894, 1990; Chardin, H., Gokani, J.P., Septier, D. et al., Structural variations of different oral basement membranes revealed by cationic dyes and detergent added to aldehyde fixative solution, Histochem. J. 24, 375–382, 1992; Agren, U.M., Tammi, R., and Tammi, M., A dot-blot assay of metabolically radiolabeled hyaluronan, Anal. Biochem. 217, 311–315, 1994; Maccari, F. and Volpi, N., Glycosaminoglycan blotting on nitrocellulose membranes treated with cetylpyridinium chloride afer agarose-gel electrophoretic separation, Electrophoresis 23, 3270–3277, 2002; Maccari, F. and Volpi, N., Direct and specific recognition of glycosaminoglycans by antibodies after their separation by agarose gel electrophoresis and blotting on cetylpyridinium chloride-treated nitrocellulose membranes, Electrophoresis 24, 1347–1352, 2003.


Hjelmeland, L.M., A nondenaturing zwitterionic detergent for membrane biochemistry: design and synthesis, Proc. Natl. Acad. Sci. USA 77, 6368–6370, 1980; Giradot, J.M. and Johnson, B.C., A new detergent for the solubilization of the vitamin K–dependent carboxylation system from liver microsomes: comparison with triton X-100, Anal. Biochem. 121, 315–320, 1982; Liscia, D.S., Alhadi, T., and Vonderhaar, B.K., Solubilization of active prolactin receptors by a nondenaturing zwitterionic detergent, J. Biol. Chem. 257, 9401–9405, 1982; Womack, M.D., Kendall, D.A., and MacDonald, R.C., Detergent effects on enzyme activity and solubilization of lipid bilayer membranes, Biochim. Biophys. Acta 733, 210–215, 1983; Klaerke, D.A. and Jorgensen, P.L., Role of Ca2+-activated K+ channel in regulation of NaCl reabsorption in thick ascending limb of Henle’s loop, Comp. Biochem. Physiol. A 90, 757–765, 1988; Kuriyama, K., Nakayasu, H., Mizutani, H. et al., Cerebral GABAB receptor: proposed mechanisms of action and purification procedures, Neurochem. Res. 18, 377–383, 1993; Koumanov, K.S., Wolf, C., and Quinn, P.J., Lipid composition of membrane domains, Subcell. Biochem. 37, 153–163, 2004.




Used for extraction of lipids, usually in combination with methanol.

Stevan, M.A. and Lyman, R.L., Investigations on extraction of rat plasma phospholipids, Proc. Soc. Exp. Biol. Med. 114, 16–20, 1963; Wells, M.A. and Dittmer, J.C., A microanalytical technique for the quantitative determination of twenty-four classes of brain lipids, Biochemistry 5, 3405–3418, 1966; Colacicco, G. and Rapaport, M.M., A simplified preparation of phosphatidyl inositol, J. Lipid. Res. 8, 513–515, 1967; Curtis, P.J., Solubility of mitochondrial membrane proteins in acidic organic solvents, Biochim. Biophys. Acta 183, 239–241, 1969; Privett, O.S., Dougherty, K.A., and Castell, J.D., Quantitative analysis of lipid classes, Am. J. Clin. Nutr. 24, 1265–1275, 1971; Claire, M., Jacotot, B., and Robert, L., Characterization of lipids associated with macromolecules of the intercellular matrix of human aorta, Connect. Tissue Res. 4, 61–71, 1976; St. John, L.C. and Bell, F.P., Extraction and fractionation of lipids from biological tissues, cells, organelles, and fluids, Biotechniques 7, 476–481, 1989; Dean, N.M. and Beaven, M.A., Methods for the analysis of inositol phosphates, Anal. Biochem. 183, 199–209, 1989; Singh, A.K. and Jiang, Y., Quantitative chromatographic analysis of inositol phospholipids and related compounds, J. Chromatog. B Biomed. Appl. 671, 255–280, 1995.


Doree, C., The occurrence and distribution of cholesterol and allied bodies in the animal kingdom, Biochem. J. 4, 72–106, 1909; Heilbron, I.M., Kamm, E.D., and Morton, R.A., The absorption spectrum of cholesterol and its biological significance with reference to vitamin D. Part I: Preliminary observations, Biochem. J. 21, 78–85, 1927; Cook, R.P., Ed., Cholesterol: Chemistry, Biochemistry, and Pathology, Academic Press, New York, 1958; Vahouny, G.V. and Treadwell, C.R., Enzymatic synthesis and hydrolysis of cholesterol esters, Methods Biochem. Anal. 16, 219–272, 1968; Heftmann, E., Steroid Biochemistry, Academic Press, New York, 1970; Nestel, P.J., Cholesterol turnover in man, Adv. Lipid Res. 8, 1–39, 1970; Dennick, R.G., The intracellular organization of cholesterol biosynthesis. A review, Steroids Lipids Res. 3, 236–256, 1972; J. Polonovski, Ed., Cholesterol Metabolism and Lipolytic Enzymes, Masson Publications, New York, 1977; Gibbons, G.F., Mitrooulos, K.A., and Myant, N.B., Biochemistry of Cholesterol, Elsevier, Amsterdam, 1982; Bittman, R., Cholesterol: Its Functions and Metabolism in Biology and Medicine, Plenum Press, New York, 1997; Oram, J.P. and Heinecke, J.W., ATP-binding cassette transporter A1: a cell cholesterol exporter that protects against cardiovascular disease, Physiol. Rev. 85, 1343–1372, 2005; Holtta-Vuori, M. and Ikonen, E., Endosomal cholesterol traffic: vesicular and non-vesicular mechanisms meet, Biochem. Soc. Trans. 34, 392–394, 2006; Cuchel, M. and Rader, D.J., Macrophage reverse cholesterol transport: key to the regression of atherosclerosis? Circulation 113, 2548–2555, 2006.


Schreiber, A.J. and Simon, F.R., Overview of clinical aspects of bile salt physiology, J. Pediatr. Gastroenterol. Nutr. 2, 337–345, 1983; Chiang, J.Y., Regulation of bile acid synthesis, Front. Biosci. 3, dl176–dl193, 1998; Cybulsky, M.I., Lichtman, A.H., Hajra, L., and Iiyama, K., Leukocyte adhesion molecules in atherogenesis, Clin. Chim. Acta 286, 207–218, 1999.


Dixon, H.B. and Perham, R.N., Reversible blocking of amino groups with citraconic anhydride, Biochem. J. 109, 312–314, 1968; Gibbons, I. and Perham, R.N., The reaction of aldolase with 2-methylmaleic anhydride, Biochem. J. 116, 843–849, 1970; Yankeelov, J.A., Jr. and Acree, D., Methylmaleic anhydride as a reversible blocking agent during specific arginine modification, Biochem. Biophys. Res. Commun. 42, 886–891, 1971; Takahashi, K., Specific modification of arginine residues in proteins with ninhydrin, J. Biochem. 80, 1173–1176, 1976; Brinegar, A.C. and Kinsella, J.E., Reversible modification of lysine in soybean proteins, using citraconic anhydride: characterization of physical and chemical changes in soy protein isolate, the 7S globulin, and lipoxygenase, J. Agric. Food Chem. 28, 818–824, 1980; Shetty, J.K. and Kensella, J.F., Ready separation of proteins from nucleoprotein complexes by reversible modification of lysine residues, Biochem. J. 191, 269–272, 1980; Yang, H. and Frey, P.A., Dimeric cluster with a single reactive amino group, Biochemistry 23, 3863–3868, 1984; Bindels, J.G., Misdom, L.W., and Hoenders, H.J., The reaction of citraconic anhydride with bovine alpha-crystallin lysine residues. Surface probing and dissociation-reassociation studies, Biochim. Biophys. Acta 828, 255–260, 1985; Al jamal, J.A., Characterization of different reactive lysines in bovine heart mitochondrial porin, Biol. Chem. 383, 1967–1970, 2002; Kadlik, V., Strohalm, M., and Kodicek, M., Citraconylation — a simple method for high protein sequence coverage in MALDI-TOF mass spectrometry, Biochem. Biophys. Res. Commun. 305, 1091–1093, 2003.


Mitchell, P., Crystallization of Congo red, Nature 165, 772–773, 1950; Helander, S., The distribution of Congo red in the tissues, Acta. Med. Scand. 138, 188–190, 1950; Hahn, N.J., The Congo red reaction in bacteria and its usefulness in the identification of rhizobia, Can. J. Microbiol. 12, 725–733, 1966; R.W. Horobin and J.A. Kiernan, Eds., Conn’s Biological Stains, 10th ed., Bios Scientific Publishers, Oxford, UK, 2002; Inouye, H. and Kirschner, D.A., Alzheimer’s beta-amyloid: insights into fibril formation and structure from Congo red binding, Subcell. Biochem. 38, 203–224, 2005; Inestrosa, N.C., Alvarez, A., Dinamarca, M.C. et al., Acetylcholinesterase-amyloid-beta-protein interaction: effect of Congo red and the role of the Wnt pathway, Curr. Alzheimer Res. 2, 301–306, 2005; Wu, X., Sun, S., Guo, C. et al., Resonance light scattering technique for the determination of proteins with Congo red and Triton X-100, Luminescence 21, 56–61, 2006; Halimi, M., Dayan-Amouyal, Y., Kariv-Inbal, Z. et al., Prion urine comprises a glycosaminoglycan-light chain IgG complex that can be stained by Congo red, J. Virol. Methods 133, 205–210, 2006; Bely, M. and Makovitzky, J., Sensitivity and specificity of Congo red staining according to Romhanyi. Comparison with Puchtler’s or Bennhold’s methods, Acta Histochem. 108, 175–180, 2006; McLaughlin, R.W., De Stigter, J.K., Sikkink, L.A. et al., The effects of sodium sulfate, glycosaminoglycans, and Congo red on the structure, stability, and amyloid formation of an immunoglobulin light-chain protein, Protein Sci. 15, 1710–1722, 2006; Cheung, S.T., Maheshwari, M.B., and Tan, C.Y., A comparative study of two Congo red stains for the detection of primary cutaneous amyloidosis, J. Am. Acad. Dermatol. 55, 363–364, 2006.


Bradford, M.M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Anal. Biochem. 72, 248–254, 1976; Saleemuddin, M., Ahmad, H., and Husain, A., A simple, rapid, and sensitive procedure for the assay of endoproteases using Coomassie Brilliant Blue G-250, Anal. Biochem. 105, 202–206, 1980; van Wilgenburg, M.G., Werkman, E.M., van Gorkom, W.H., and Soons, J.B., Criticism of the use of Coomassie Brilliant Blue G-250 for the quantitative determination of proteins, J.Clin. Chem.Clin. Biochem. 19, 301–304, 1981; Mattoo, R.L., Ishaq, M., and Saleemuddin, M., Protein assay by Coomassie Brilliant Blue G-250-binding method is unsuitable for plant tissues rich in phenols and phenolases, Anal. Biochem. 163, 376–384, 1987; Lott, J.A., Stephan, V.A., and Pritchard, K.A., Jr., Evaluation of the Coomassie Brilliant Blue G-250 method for urinary proteins, Clin. Chem. 29, 1946–1950, 1983; Fanger, B.O., Adaptation of the Bradford protein assay to membrane-bound proteins by solubilizing in glucopyranoside detergents, Anal. Biochem. 162, 11–17, 1987; Marshall, T. and Williams, K.M., Recovery of proteins by Coomassie Brilliant Blue precipitation prior to electrophoresis, Electrophoresis 13, 887–888, 1992; Sapan, C.V., Lundblad, R.L., and Price, N.C., Colorimetric protein assay techniques, Biotechnol. Appl. Biochem. 29, 99–108, 1999.


Vesterberg, O., Hansen, L., and Sjosten, A., Staining of proteins after isoelectric focusing in gels by a new procedure, Biochim. Biophys. Acta 491, 160–166, 1977; Micko, S. and Schlaepfer, W.W., Metachromasy of peripheral nerve collagen on polyacrylamide gels stained with Coomassie Brilliant Blue R-250, Anal. Biochem. 88, 566–572, 1978; Osset, M., Pinol, M., Fallon, M.J. et al., Interference of the carbohydrate moiety in Coomassie Brilliant Blue R-250 protein staining, Electrophoresis 10, 271–273, 1989; Pryor, J.L., Xu, W., and Hamilton, D.W., Immunodetection after complete destaining of Coomassie blue-stained proteins on immobilon-PVDF, Anal. Biochem. 202, 100–104, 1992; Metkar, S.S., Mahajan, S.K., and Sainis, J.K., Modified procedure for nonspecific protein staining on nitrocellulose paper using Coomassie Brilliant Blue R-250, Anal. Biochem. 227, 389–391, 1995; Kundu, S.K., Robey, W.G., Nabors, P. et al., Purification of commercial Coomassie Brilliant Blue R-250 and characterization of the chromogenic fractions, Anal. Biochem. 235, 134–140, 1996; Choi, J.K., Yoon, S.H., Hong, H.Y. et al., A modified Coomassie blue staining of proteins in polyacrylamide gels with Bismark brown R, Anal. Biochem. 236, 82–84, 1996; Moritz, R.L., Eddes, J.S., Reid, G.E., and Simpson, R.J., S-pyridylethylation of intact polyacrylamide gels and in situ digestion of electrophoretically separated proteins: a rapid mass spectrometric method for identifying cysteine-containing peptides, Electrophoresis 17, 907–917, 1996; Choi, J.K. and Yoo, G.S., Fast protein staining in sodium dodecyl sulfate polyacrylamide gel using counter ion-dyes, Coomassie Brilliant Blue R-250, and neutral red, Arch. Pharm. Res. 25, 704–708, 2002; Bonar, E., Dubin, A., Bierczynska-Krzysik, A. et al., Identification of major cellular proteins synthesized in response to interleukin-1 and interleukin-6 in human hepatoma HepG2 cells, Cytokine 33, 111–117, 2006.


Tonge, R., Shaw, J., Middleton, B. et al., Validation and development of fluorescence two-dimensional differential gel electrophoresis proteomics technology, Proteomics 1, 377–396, 2001; Chan, H.L., Gharbi, S., Gaffney, P.R. et al., Proteomic analysis of redox- and ErbB2-dependent changes in mammary luminal epithelial cells using cysteine- and lysine-labeling two-dimensional difference gel electrophoresis, Proteomics 5, 2908–2926, 2005; Misek, D.E., Kuick, R., Wang, H. et al., A wide range of protein isoforms in serum and plasma uncovered by a quantitative intact protein analysis system, Proteomics 5, 3343–3352, 2005; Doutette, P., Navet, R., Gerkens, P. et al., Steatosis-induced proteomic changes in liver mitochondria evidenced by two-dimensional differential in-gel electrophoresis, J. Proteome Res. 4, 2024–2031, 2005.


Brismar, H. and Ulfake, B., Fluorescence lifetime measurements in confocal microscopy of neurons labeled with multiple fluorophores, Nat. Biotechnol. 15, 373–377, 1997; Strohmaier, A.R., Porwol, T., Acker, H., and Spiess, E., Tomography of cells by confocal laser scanning microscopy and computer-assisted three-dimensional image reconstruction: localization of cathepsin B in tumor cells penetrating collagen gels in vitro, J. Histochem. Cytochem. 45, 975–983, 1997; Alexandre, I., Hamels, S., Dufour, S. et al., Colorimetric silver detection of DNA microarrays, Anal. Biochem. 295, 1–8, 2001; Shaw, J., Rowlinson, R., Nickson, J. et al., Evaluation of saturation labeling two-dimensional difference gel electrophoresis fluorescent dyes, Proteomics 3, 1181–1195, 2003.


Uchihara, T., Nakamura, A., Nagaoka, U. et al., Dual enhancement of double immunofluorescent signals by CARD: participation of ubiquitin during formation of neurofibrillary tangles, Histochem. Cell Biol. 114, 447–451, 2000; Duthie, R.S., Kalve, I.M., Samols, S.B. et al., Novel cyanine dye-based dideoxynucleoside triphosphates for DNA sequencing, Bioconjug. Chem. 13, 699–706, 2002; Graves, E.E., Yessayan., D., Turner, G. et al., Validation of in vivo fluorochrome concentrations measured using fluorescence molecular tomography, J. Biomed. Opt. 10, 44019, 2005; Lapeyre, M., Leprince, J., Massonneau, M. et al., Aryldithioethyloxycarbonyl (Ardec): a new family of amine-protecting groups removable under mild reducing conditions and their applications to peptide synthesis, Chemistry 12, 3655–3671, 2006; Tang, X., Morris, S.L., Langone, J.J., and Bockstahler, L.E., Simple and effective method for generating single-stranded DNA targets and probes, Biotechniques 40, 759–763, 2006.



Gobom, J., Schuerenberg, M., Mueller, M. et al., α-cyano-4-hydroxycinnamic acid affinity sample preparation. A protocol for MALDI-MS peptide analysis in proteomics, Anal. Chem. 73, 434–438, 2001; Zhu, X. and Papayannopoulos, I.A., Improvement in the detection of low concentration protein digests on a MALDI TOF/TOF workstation by reducing α-cyano-4-hydroxycinnamic acid adduct ions, J. Biomol. Tech. 14, 298–307, 2003; Neubert, H., Halket, J.M., Fernandez Ocana, M., and Patel, R.K., MALDI post-source decay and LIFT-TOF/TOF investigation of α-cyano-4-hydroxycinnamic acid cluster interferences, J. Am. Soc. Mass Spectrom. 15, 336–343, 2004; Kobayashi, T., Kawai, H., Suzuki, T. et al., Improved sensitivity for insulin in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry by premixing α-cyano-4-hydroxycinnamic acid with transferrin, Rapid Commun. Mass Spectrom. 18, 1156–1160, 2004; Pshenichnyuk, S.A. and Asfandiarov, N.L., The role of free electrons in MALDI: electron capture by molecules of α-cyano-4-hydroxycinnamic acid, Eur. J. Mass Spectrom. 10, 477–486, 2004; Bogan, M.J., Bakhoum, S.F., and Agnes, G.R., Promotion of α-cyano-4-hydroxycinnamic acid and peptide cocrystallization within levitated droplets with net charge, J. Am. Soc. Mass Spectrom. 16, 254–262, 2005. As enzyme inhibitor: Clarke, P.D., Clift, D.L., Dooledeniya, M. et al., Effects of α-cyano-4-hydroxycinnamic acid on fatigue and recovery of isolated mouse muscle, J. Muscle Res. Cell Motil. 16, 611–617, 1995; Del Prete, E., Lutz, T.A., and Scharrer, E., Inhibition of glucose oxidation by α-cyano-4-hydroxycinnamic acid stimulates feeding in rats, Physiol. Behav. 80, 489–498, 2004; Briski, K.P. and Patil, G.D., Induction of Fox immunoreactivity labeling in rat forebrain metabolic loci by caudal fourth ventricular infusion of the monocarboxylate transporter inhibitor, α-cyano-4-hydroxycinnamic acid, Neuroendocrinology 82, 49–57, 2005.


Ghenbot, G., Emge, T., and Day, R.A., Identification of the sites of modification of bovi carbonic anhydrase II (BCA II) by the salt bridge reagent cyanogen, C2N2, Biochim. Biophys. Acta 1161, 59–65, 1993; Karagozler, A.A., Ghenbot, G., and Day, R.A., Cyanogen as a selective probe for carbonic anhydrase hydrolase, Biopolymers 33, 687–692, 1993; Winters, M.S. and Day, R.A., Identification of amino acid residues participating in intermolecular salt bridges between self-associating proteins, Anal. Biochem. 309, 48–59, 2002; Winters, M.S. and Day, R.A., Detecting protein–protein interactions in the intact cell of Bacillus subtilis(ATCC 6633), J. Bacteriol. 185, 4268–4275, 2003.


Hofmann, T., The purification and properties of fragments of trypsinogen obtained by cyanogen bromide cleavage, Biochemistry 3, 356–364, 1964; Chu, R.C. and Yasunobu, K.T., The reaction of cyanogen bromide and N-bromosuccinimide with some cytochromes C, Biochim. Biophys. Acta 89, 148–149, 1964; Inglis, A.S. and Edman, P., Mechanism of cyanogen bromide reaction with methionine in peptides and proteins. I. Formation of imidate and methyl thiocyanate, Anal. Biochem. 37, 73–80, 1970; Kagedal, L. and Akerstrom, S., Binding of covalent proteins to polysaccharides by cyanogen bromide and organic cyanates. I. Preparation of soluble glycine-, insulin- and ampicillin-dextran, Acta Chem. Scand. 25, 1855–1899, 1971; Sipe, J.D. and Schaefer, F.V., Preparation of solid-phase immunosorbents by coupling human serum proteins to cyanogen bromide–activated agarose, Appl. Microbiol. 25, 880–884, 1973; March, S.C., Parikh, I., and Cuatrecasas, P., A simplified method for cyanogen bromide activation of agarose for affinity chromatography, Anal. Biochem. 60, 149–152, 1974; Boulware, D.W., Goldsworthy, P.D., Nardella, F.A., and Mannik, M., Cyanogen bromide cleaves Fc fragments of pooled human IgG at both methionine and tryptophan residues, Mol. Immunol. 22, 1317–1322, 1985; Jaggi, K.S. and Gangal, S.V., Monitoring of active groups of cyanogen bromide-activated paper discs used as allergosorbent, Int. Arch. Allergy Appl. Immunol. 89, 311–313, 1989; Villa, S., De Fazio, G., and Canosi, U., Cyanogen bromide cleavage at methionine residues of polypeptides containing disulfide bonds, Anal. Biochem. 177, 161–164, 1989; Luo, K.X., Hurley, T.R., and Sefton, B.M., Cyanogen bromide cleavage and proteolytic peptide mapping of proteins immobilized to membranes, Methods Enzymol. 201, 149–152, 1991; Jennissen, H.P., Cyanogen bromide and tresyl chloride chemistry revisited: the special reactivity of agarose as a chromatographic and biomaterial support for immobilizing novel chemical groups, J. Mol. Recognit. 8, 116–124, 1995; Kaiser, R. and Metzka, L., Enhancement of cyanogen bromide cleavage yields for methionyl-serine and methionyl-threonine peptide bonds, Anal. Biochem. 266, 1–8, 1999; Kraft, P., Mills, J., and Dratz, E., Mass spectrometric analysis of cyanogen bromide fragments of integral membrane proteins at the picomole level: application to rhodopsin, Anal. Biochem. 292, 76–86, 2001; Kuhn, K., Thompson, A., Prinz, T. et al., Isolation of N-terminal protein sequence tags from cyanogen bromide-cleaved proteins as a novel approach to investigate hydrophobic proteins, J. Proteome Res. 2, 598–609, 2003; Macmillan, D. and Arham, L., Cyanogen bromide cleavage generates fragments suitable for expressed protein and glycoprotein ligation, J. Am. Chem. Soc. 126, 9530–9531, 2004; Lei, H., Minear, R.A., and Marinas, B.J., Cyanogen bromide formation from the reactions of monobromamine and dibromamine with cyanide ions, Environ. Sci. Technol. 40, 2559–2564, 2006.


Gray, B.M., ELISA methodology for polysaccharide antigens: protein coupling of polysaccharides for adsorption to plastic tubes, J. Immunol. Methods 28, 187–192, 1979: Horak, D., Rittich, B., Safar, J. et al., Properties of RNase A immobilized on magnetic poly(2-hydroxyethyl methacrylate) microspheres, Biotechnol. Prog. 17, 447–452, 2001; Lee, P.H., Sawan, S.P., Modrusan, Z. et al., An efficient binding chemistry for glass polynucleotide microarrays, Bioconjug. Chem. 13, 97–103, 2002; Steinberg, G., Stromsborg, K., Thomas, L. et al., Strategies for covalent attachment of DNA to beads, Biopolymers 73, 597–605, 2004; Abuknesha, R.A., Luk, C.Y., Griffith, H.H. et al., Efficient labeling of antibodies with horseradish peroxidase using cyanuric chloride, J. Immunol. Methods 306, 211–217, 2005.


Tandon, S.K. and Singh, J., Removal of manganese by chelating agents from brain and liver of manganese, Toxicology 5, 237–241, 1975; Hazell, A.S., Normandin, L., Norenberg, M.D., Kennedy, G., and Yi, J.H., Alzheimer type II astrocyte changes following sub-acute exposure to manganese, Neurosci. Lett., 396, 167–171, 2006; Hassler, C.S. and Twiss, M.R., Bioavailability of iron sensed by a phytoplanktonic Fe-bioreporter, Environ. Sci. Tech. 40, 2544–2551, 2006.

Dansyl Chloride

5-(dimethylamino)-1-naphthalenesulfonyl Chloride


Fluorescent label for proteins; amino acid analysis.

Hill, R.D. and Laing, R.R., Specific reaction of dansyl chloride with one lysine residue in rennin, Biochim. Biophys. Acta 132, 188–190, 1967; Chen, R.F., Fluorescent protein-dye conjugates. I. Heterogeneity of sites on serum albumin labeled by dansyl chloride, Arch. Biochem. Biophys. 128, 163–175, 1968; Chen, R.F., Dansyl-labeled protein modified with dansyl chloride: activity effects and fluorescence properties, Anal. Biochem. 25, 412–416, 1968; Brown, C.S. and Cunningham, L.W., Reaction of reactive sulfydryl groups of creatine kinase with dansyl chloride, Biochemistry 9, 3878–3885, 1970; Hsieh, W.T. and Matthews, K.S., Lactose repressor protein modified with dansyl chloride: activity effects and fluorescence properties, Biochemistry 34, 3043–3049, 1985; Scouten, W.H., van den Tweel, W., Kranenburg, H., and Dekker, M., Colored sulfonyl chloride as an activated agent for hydroxylic matrices, Methods Enzymol. 135, 79–84, 1987; Martin, M.A., Lin, B., Del Castillo, B., The use of fluorescent probes in pharmaceutical analysis, J. Pharm. Biomed. Anal. 6, 573–583, 1988; Walker, J.M., The dansyl method for identifying N-terminal amino acids, Methods Mol. Biol. 32, 321–328, 1994; Walker, J.M., The dansyl-Edman method for peptide sequencing, Methods Mol. Biol. 32, 329–334, 1994; Pin, S. and Royer, C.A., High-pressure fluorescence methods for observing subunit dissociation in hemoglobin, Methods Enzymol. 323, 42–55, 1994; Rangarajan, B., Coons, L.S., and Scarnton, A.B., Characterization of hydrogels using luminescence spectroscopy, Biomaterials 17, 649–661, 1996; Kang, X., Xiao, J., Huang, X., and Gu, X., Optimization of dansyl derivatization and chromatographic conditions in the determination of neuroactive amino acids of biological samples, Clin. Chim. Acta 366, 352–356, 2006.


Chau, A.S. and Terry, K., Analysis of pesticides by chemical derivatization. I. A new procedure for the formation of 2-chloroethyl esters of ten herbicidal acids, J. Assoc. Off. Anal. Chem. 58, 1294–1301, 1975; Patel, L. and Kaback, H.R., The role of the carbodiimide-reactive component of the adenosine-5′-triphosphatase complex in the proton permeability of Escherichia coli membrane vesicles, Biochemistry 15, 2741–2746, 1976; Esch, F.S., Bohlen, P., Otsuka, A.S. et al., Inactivation of the bovine mitochondrial F1-ATPase with dicyclohexyl[14C]carbodiimide leads to the modification of a specific glutamic acid residue in the beta subunit, J. Biol. Chem. 256, 9084–9089, 1981; Hsu, C.M. and Rosen, B.P., Characterization of the catalytic subunit of an anion pump, J. Biol. Chem. 264, 17349–17354, 1989; Gurdag, S., Khandare, J., Stapels, S. et al., Activity of dendrimer-methotrexate conjugates on methotrexate-sensitive and -resistant cell lines, Bioconjug. Chem. 17, 275–283, 2006; Vgenopoulou, I., Gemperli, A.C., and Steuber, J., Specific modification of a Na+ binding site in NADH: quinone oxidoreductase from Klebsiella pneumoniae with dicyclohexylcarbodiimide, J. Bacteriol. 188, 3264–3272, 2006; Ferguson, S.A., Keis, S., and Cook, G.M., Biochemical and molecular characterization of a Na+-translocating F1Fo-ATPase from the thermophilic bacterium Clostridium paradoxum, J. Bacteriol. 188, 5045–5054, 2006.

Deoxycholic Acid

Desoxycholic Acid


Detergent, nanoparticles.

Akare, S. and Martinez, J.D., Bile acid-induced hydrophobicity-dependent membrane alterations, Biochim. Biophys. Acta 1735, 59–67, 2005; Chae, S.Y., Son, S., Lee, M. et al., Deoxycholic acid-conjugated chitosan oligosaccharide nanoparticles for efficient gene carrier, J. Control. Release 109, 330–344, 2005; Dall’Agnol, M., Bernstein, C., Bernstein, H. et al., Identification of S-nitrosylated proteins after chronic exposure of colon epithelial cells to deoxycholate, Proteomics 6, 1654–1662, 2006; Dotis, J., Simitsopoulou, M., Dalakiouridou, M. et al. Effects of lipid formulations of amphotericin B on activity of human monocytes against Aspirgillus fumigatus, Antimicrob. Agents Chemother. 128, 3490–3491, 2006; Darragh, J., Hunter, M., Pohler, E. et al., The calcium-binding domain of the stress protein SEP53 is required for survival in response to deoxycholic acid-mediated injury, FEBS J. 273, 1930–1947, 2006.

Deuterium Oxide

“Heavy Water”


Structural studies in proteins, enzyme kinetics; in vivo studies of metabolic flux.

Cohen, A.H., Wilkinson, R.R., and Fisher, H.F., Location of deuterium oxide solvent isotope effects in the glutamate dehydrogenase reaction, J. Biol. Chem. 250, 5343–5246, 1975; Rosenberry, T.L., Catalysis by acetylcholinesterase: evidence that the rate-limiting step for acylation with certain substrates precedes general acid-base catalysis, Proc. Natl. Acad. Sci. USA 72, 3834–3838, 1975; Viggiano, G., Ho, N.T., and Ho, C., Proton nuclear magnetic resonance and biochemical studies of oxygenation of human adult hemoglobin in deuterium oxide, Biochemistry 18, 5238–5247, 1979; Bonnete, F., Madern, D., and Zaccai, G., Stability against denaturation mechanisms in halophilic malate dehydrogenase “adapt” to solvent conditions, J. Mol. Biol. 244, 436–447, 1994; Thompson, J.F., Bush, K.J., and Nance, S.L., Pancreatic lipase activity in deuterium oxide, Proc. Soc. Exp. Biol. Med. 122, 502–505, 1996; Dufner, D. and Previs, S.F., Measuring in vivo metabolism using heavy water, Curr. Opin. Clin. Nutr. Metab. Care 6, 511–517, 2003; O’Donnell, A.H., Yao, X., and Byers, L.D., Solvent isotope effects on alpha-glucosidase, Biochem. Biophys. Acta 1703, 63–67, 2004; Hellerstein, M.K. and Murphy, E., Stable isotope-mass spectrometric measurements of molecular fluxes in vivo: emerging applications in drug development, Curr. Opin. Mol. Ther. 6, 249–264, 2004; Mazon, H., Marcillat, O., Forest, E., and Vial, C., Local dynamics measured by hydrogen/deuterium exchange and mass spectrometry of the creatine kinase digested by two proteases, Biochimie 87, 1101–1110, 2005; Carmieli, R., Papo, N., Zimmerman, H. et al., Utilizing ESEEM spectrscopy to locate the position of specific regions of membrane-active peptides within model membranes, Biophys. J. 90, 492–505, 2006.


Baker, B.R., Factors in the design of active-site-directed irreversible inhibitors, J. Pharm. Sci. 53, 347–364, 1964; Dixon, G.H. and Schachter, H., The chemical modification of chymotrypsin, Can. J. Biochem. Physiol. 42, 695–714, 1964; Singer, S.J., Covalent labeling active site, Adv. Protein Chem. 22, 1–54, 1967; Kassell, B. and Kay, J., Zymogens of proteolytic enzymes, Science 180, 1022–1027, 1973; Fujino, T., Watanabe, K., Beppu, M. et al., Identification of oxidized protein hydrolase of human erythrocytes as acylpeptide hydrolase, Biochim. Biophys. Acta 1478, 102–112, 2000; Manco, G., Camardello, L., Febbraio, F. et al., Homology modeling and identification of serine 160 as nucleophile as the active site in a thermostable carboxylesterase from the archeon Archaeoglobus fulgidus, Protein Eng. 13, 197–200, 2000; Gopal, S., Rastogi, V., Ashman, W., and Mulbry, W., Mutagenesis of organophosphorous hydrolase to enhance hydrolysis of the nerve agent VX, Biochem. Biophys. Res. Commun. 279, 516–519, 2000; Yeung, D.T., Lenz, D.E., and Cerasoli, D.M., Analysis of active-site amino acid residues of human serum paraoxanse using competitive substrates, FEBS J. 272, 2225–2230, 2005; D’Souza, C.A., Wood, D.D., She, Y.M., and Moscarello, M.A., Autocatalytic cleavage of myelin basic protein: an alternative to molecular mimicry, Biochemistry 44, 12905–12913, 2005.


Bouillon, R., Kerkhove, P.V., and De Moor, P., Measurement of 25-hydroxyvitamin D3 in serum, Clin. Chem. 22, 364–368, 1976; Redhwi, A.A., Anderson, D.C., and Smith, G.N., A simple method for the isolation of vitamin D metabolites from plasma extracts, Steroids 39, 149–154, 1982; Scholtz, R., Wackett, L.P., Egli, C. et al., Dichloromethane dehalogenase with improved catalytic activity isolated form a fast-growing dichloromethane-utilizing bacterium, J. Bacteriol. 170, 5698–5704, 1988; Russo, M.V., Goretti, G., and Liberti, A., Direct headspace gas chromatographic determination of dichloromethane in decaffeinated green and roasted coffee, J. Chromatog. 465, 429–433, 1989; Shimizu, M., Kamchi, S., Nishii, Y., and Yamada, S., Synthesis of a reagent for fluorescence-labeling of vitamin D and its use in assaying vitamin D metabolites, Anal. Biochem. 194, 77–81, 1991; Rodriguez-Palmero, M., de la Presa-Owens, S., Castellote-Bargallo, A.I. et al., Determination of sterol content in different food samples by capillary gas chromatography, J. Chromatog. A 672, 267–272, 1994; Raghuvanshi, R.S., Goyal, S., Singh, O., and Panda, A.K., Stabilization of dichloromethane-induced protein denaturation during microencapsulation, Pharm. Dev. Technol. 3, 269–276, 1998; El Jaber-Vazdekis, N., Gutierrez-Nicolas, F., Ravelo, A.G., and Zarate, R., Studies on tropane alkaloid extraction by volatile organic solvents: dichloromethane vs. chloroform, Phytochem. Anal. 17, 107–113, 2006.


Matsuba, Y. and Takahashi, Y., Spectrophotometric determination of copper with N,N,N,N′-tetraethylthiuram disulfide and an application of this method for studies on subcellular distribution of copper in rat brains, Anal. Biochem. 36, 182–191, 1970, Koutensky, J., Eybl, V., Koutenska, M. et al., Influence of sodium diethyldithiocarbamate on the toxicity and distribution of copper in mice, Eur. J. Pharmacol. 14, 389–392, 1971; Xu, H. and Mitchell, C.L., Chelation of zinc by diethyldithiocarbamate facilitates bursting induced by mixed antidromic plus orthodromic activation of mossy fibers in hippocampal slices, Brain Res. 624, 162–170, 1993; Liu, J., Shigenaga, M.K., Yan, L.J. et al., Antioxidant activity of diethyldithiocarbamate, Free Radic. Res. 24, 461–472, 1996; Zhang, Y., Wade, K.L., Prestera, T., and Talalav, P., Quantitative determination of isothiocyanates, dithiocarbamates, carbon disulfide, and related thiocarbonyl compounds by cyclocondensation with 1,2-benzenedithiol, Anal. Biochem. 239, 160–167, 1996; Shoener, D.F., Olsen, M.A., Cummings, P.G., and Basic, C., Electrospray ionization of neutral metal dithiocarbamate complexes using in-source oxidation, J. Mass Spectrom. 34, 1069–1078, 1999; Turner, B.J., Lopes, E.C., and Cheema, S.S., Inducible superoxide dismutase 1 aggregation in transgenic amyotrophic lateral sclerosis mouse fibroblasts, J. Cell Biochem. 91, 1074–1084, 2004; Xu, K.Y. and Kuppusamy, P., Dual effects of copper-zinc superoxide dismutase, Biochem. Biophys. Res. Commun. 336, 1190–1193, 2005; Jiang, X., Sun, S., Liang, A. et al., Luminescence properties of metal(II)-diethyldithiocarbamate chelate complex particles and its analytical application, J. Fluoresc. 15, 859–864, 2005; Wang, J.S. and Chiu, K.H., Mass balance of metal species in supercritical fluid extraction using sodium diethyldithiocarbamate and dibuylammonium dibutyldithiocarbamate, Anal. Sci. 22, 363–369, 2006.


Wolf, B., Lesnaw, J.A., and Reichmann, M.E., A mechanism of the irreversible inactivation of bovine pancreatic ribonuclease by diethylpyrocarbonate. A general reaction of diethylpyrocarbonate with proteins, Eur. J. Biochem. 13, 519–525, 1970; Splittstoesser, D.F. and Wilkison, M., Some factors affecting the activity of diethylpyrocarbonate as a sterilant, Appl. Microbiol. 25, 853–857, 1973; Fedorcsak, I., Ehrenberg, L., and Solymosy, F., Diethylpyrocarbonate does not degrade RNA, Biochem. Biophys. Res. Commun. 65, 490–496, 1975; Berger, S.L., Diethylpyrocarbonate: an examination of its properties in buffered solutions with a new assay technique, Anal. Biochem. 67, 428–437, 1975; Lloyd, A.G. and Drake, J.J., Problems posed by essential food preservatives, Br. Med. Bull. 31, 214–219, 1975; Ehrenberg, L., Fedorcsak, I., and Solymosy, F., Diethylpyrocarbonate in nucleic acid research, Prog. Nucleic Acid Res. Mol. Biol. 16, 189–262, 1976; Saluz, H.P. and Jost, J.P., Approaches to characterize protein–DNA interactions in vivo, Crit. Rev. Eurkaryot. Gene Expr. 3, 1–29, 1993; Bailly, C. and Waring, M.J., Diethylpyrocarbonate and osmium tetroxide as probes for drug-induced changes in DNA conformation in vitro, Methods Mol. Biol. 90, 51–59, 1997; Mabic, S. and Kano, I., Impact of purified water quality on molecular biology experiments, Clin. Chem. Lab. Med. 41, 486–491, 2003; Colleluori, D.M., Reczkowski, R.S., Emig, F.A. et al., Probing the role of the hyper-reactive histidine residue of argininase, Arch. Biochem. Biophys. 444, 15–26, 2005; Wu, S.N. and Chang, H.D., Diethylpyrocarbonate, a histidine-modifying agent, directly stimulates activity of ATP-sensitive potassium channels in pituitary GH(3) cells, Biochem. Pharmacol. 71, 615–623, 2006.


Bulmer, D., Dimedone as an aldehyde-blocking reagent to facilitate the histochemical determination of glycogen, Stain Technol. 34, 95–98, 1959; Sawicki, E. and Carnes, R.A., Spectrophotofluorimetric determination of aldehydes with dimedone and other reagents, Mikrochem. Acta 1, 95–98, 1968; Benitez, L.V. and Allison, W.S., The inactivation of the acyl phosphatase activity catalyzed by the sufenic acid form of glyceraldehyde 3-phosphate dehydrogenase by dimedone and olifins, J. Biol. Chem. 249, 6234–6243, 1974; Huszti, Z. and Tyihak, E., Formation of formaldehyde from S-adenosylL-[methyl-3H]methionine during enzymic transmethylation of histamine, FEBS Lett. 209, 362–366, 1986; Sardi, E. and Tyihak, E., Sample determination of formaldehyde in dimedone adduct form in biological samples by high-performance liquid chromatography, Biomed. Chromatog. 8, 313–314, 1994; Demaster, A.G., Quast, B.J., Redfern, B., and Nagasawa, H.T., Reaction of nitric oxide with the free sulfydryl group of human serum albumin yields a sulfenic acid and nitrous oxide, Biochemistry 34, 14494–14949, 1995; Rozylo, T.K., Siembida, R., and Tyihak, E., Measurement of formaldehyde as dimedone adduct and potential formaldehyde precursors in hard tissues of human teeth by overpressurized layer chromatography, Biomed. Chromatog. 13, 513–515, 1999; Percival, M.D., Ouellet, M., Campagnolo, C. et al., Inhibition of cathepsin K by nitric oxide donors: evidence for the formation of mixed disulfides and a sulfenic acid, Biochemistry 38, 13574–13583, 1999; Carballal, S., Radi, R., Kirk, M.C. et al., Sulfenic acid formation in human serum albumin by hydrogen peroxide and peroxynitrite, Biochemistry 42, 9906–9914, 2003; Poole, L.B., Zeng, B.-B., Knaggs, S.A., Yakuba, M., and King, S.B., Synthesis of chemical probes to map sulfenic acid modifications on proteins, Bioconjugate Chem. 16, 1624–1628, 2005; Kaiserov, K., Srivastava, S., Hoetker, J.D. et al., Redox activation of aldose reductase in the ischemic heart, J. Biol. Chem. 281, 15110–15120, 2006.


Eliezer, N. and Silberberg, A., Structure of branched poly-alpha-amino acids in dimethylformamide. I. Light scattering, Biopolymers 5, 95–104, 1967; Bonner, O.D., Bednarek, J.M., and Arisman, R.K., Heat capacities of ureas and water in water and dimethylformamide, J. Am. Chem. Soc. 99, 2898–2902, 1977; Sasson, S. and Notides, A.C., The effects of dimethylformamide on the interaction of the estrogen receptor with estradiol, J. Steroid Biochem. 29, 491–495, 1988; Jeffers, R.J., Feng, R.Q., Fowlkes, J.B. et al., Dimethylformamide as an enhancer of cavitation-induced cell lysis in vitro, J. Acoust. Soc. Am. 97, 669–676, 1995; You, L. and Arnold, F.H., Directed evolution of subtilisin E in Bacillus subtilis to enhance total activity in aqueous dimethylformamide, Protein Eng. 9, 77–83, 1996; Szabo, P.T. and Kele, Z., Electrospray mass spectrometry of hydrophobic compounds using dimethyl sulfoxide and dimethylformamide, Rapid Commun. Mass Spectrom. 15, 2415–2419, 2001; Nishida, Y., Shingu, Y., Dohi, H., and Kobayashi, K., One-pot alpha-glycosylation method using Appel agents in N,N-dimethylformamide, Org. Lett. 5, 2377–2380, 2003; Shingu, Y., Miyachi, A., Miura, Y. et al., One-pot alpha-glycosylation pathway via the generation in situ of alpha-glycopyranosyl imidates I N,N-dimethylformamide, Carbohydr. Res. 340, 2236–2244, 2005; Porras, S.P. and Kenndler, E., Capillary electrophoresis in N,N-dimethylformamide, Electrophoresis 26, 3279–3291, 2005; Wei, Q., Zhang, H., Duan, C. et al., High sensitive fluorophotometric determination of nucleic acids with pyronine G sensitized by N,N-dimethylformamide, Ann. Chim. 96, 273–284, 2006.


Davies, G.E. and Stark, G.R., Use of dimethyl suberimidate, a crosslinking reagent, in studying the subunit structure of oligomeric proteins, Proc. Natl. Acad. Sci. USA 66, 651–656, 1970; Hassell, J. and Hand, A.R., Tissue fixation with diimidoesters as an alternative to aldehydes. I. Comparison of crosslinking and ultrastructure obtained with dimethylsuberimidate and glutaraldehyde, J. Histochem. Cytochem. 22, 223–229, 1974; Thomas, J.O., Chemical crosslinking of histones, Methods Enzymol. 170, 549–571, 1989; Roth, M.R., Avery, R.B., and Welti, R., Crosslinking of phosphatidylethanolamine neighbors with dimethylsuberimidate is sensitive to the lipid phase, Biochim. Biophys. Acta 986, 217–224, 1989; Redl, B., Walleczek, J., Soffler-Meilicke, M., and Stoffler, G., Immunoblotting analysis of protein–protein crosslinks within the 50S ribosomal subunit of Escherichia coli. A study using dimethylsuberimidate as crosslinking reagent, Eur. J. Biochem. 181, 351–256, 1989; Konig, S., Hubner, G., and Schellenberger, A., Crosslinking of pyruvate decarboxylase-characterization of the native and substrate-activated enzyme states, Biomed. Biochim. Acta 49, 465–471, 1990; Chen, J.C., von Lintig, F.C., Jones, S.B. et al., High-efficiency solid-phase capture using glass beads bonded to microcentrifuge tubes: immunoprecipitation of proteins from cell extracts and assessment of ras activation, Anal. Biochem. 302, 298–304, 2002; Dufes, C., Muller, J.M., Couet, W. et al., Anticancer drug delivery with transferrin-targeted polymeric chitosan vesicles, Pharm. Res. 21, 101–107, 2004; Levchenko, V. and Jackson, V., Histone release during transcription: NAP1 forms a complex with H2A and H2B and facilitates a topologically dependent release of H3 and H4 from the nucleosome, Biochemistry 43, 2358–2372, 2004; Jastrzebska, M., Barwinski, B., Mroz, I. et al., Atomic force microscopy investigation of chemically stabilized pericardium tissue, Eur. Phys. J. E 16, 381–388, 2005.


Nielsen, P.E., In vivo footprinting: studies of protein–DNA interactions in gene regulation, Bioessay 11, 152–155, 1989; Saluz, H.P. and Jost, J.P., Approaches to characterize protein–DNA interactions in vivo, Crit. Rev. Eurkaryot. Gene Expr. 3, 1–29, 1993; Saluz, H.P. and Jost, J.P., In vivo DNA footprinting by linear amplification, Methods Mol. Biol. 31, 317–329, 1994; Paul, A.L. and Ferl, R.J., In vivo footprinting of protein–DNA interactions, Methods Cell Biol. 49, 391–400, 1995; Gregory, P.D., Barbaric, S., and Horz, W., Analyzing chromatin structure and transcription factor binding in yeast, Methods 15, 295–302, 1998; Simpson, R.T., In vivo to analyze chromatin structrure, Curr. Opin. Genet. Dev. 9, 225–229, 1999; Nawrocki, A.R., Goldring, C.E., Kostadinova, R.M. et al., In vivo footprinting of the human 11β-hydroxysteroid dehydrogenase type 2 promoter: evidence for cell-specific regulation by Sp1 and Sp3, J. Biol. Chem. 277, 14647–14656, 2002; McGarry, K.C., Ryan, V.T., Grimwade, J.E., and Leonard, A.C., Two discriminatory binding sites in the Escherichia coli replication origin are required for DNA stand opening by initiator DnaA-ATP, Proc. Natl. Acad. Sci. USA 101, 2811–2816, 2004; Kellersberger, K.A., Yu, E., Kruppa, G.H. et al., Two-down characterization of nucleic acids modified by structural probes using high-resolution tandem mass spectrometry and automated data interpretation, Anal. Chem. 76, 2438–2445, 2004; Matthews, D.H., Disney, M.D., Childs, J.L. et al., Incorporating chemical modification constraints into a dynamic programming algorithm for prediction of RNA secondary structure, Proc. Natl. Acad. Sci. USA 101, 7287–7292, 2004; Forstemann, K. and Lingner, J., Telomerase limits the extent of base pairing between template RNA and temomeric DNA, EMBO Rep. 6, 361–366, 2005; Kore, A.R. and Parmar, G., An industrial process for selective synthesis of 7-methyl guanosine 5′-diphosphate: versatile synthon for synthesis of mRNA cap analogues, Nucleosides Nucleotides Nucleic Acids 25, 337–340, 2006.


Sideri, C.N. and Osol, A., A note on the purification of dioxane for use in preparing nonaqueous titrants, J. Am. Pharm. Am. Pharm. Assoc. 42, 586, 1953; Martel, R.W. and Kraus, C.A., The association of ions in dioxane-water mixtures at 25 degrees, Proc. Natl. Acad. Sci. USA 41, 9–20, 1955; Mercier, P.L. and Kraus, C.A, The ion-pair equilibrium of electrolyte solutions in dioxane-water mixtures, Proc. Natl. Acad. Sci. USA 41, 1033–1041, 1995; Inagami, T., and Sturtevant, J.M., The trypsin-catalyzed hydrolysis of benzoyl-L-arginine ethyl ester. I. The kinetics in dioxane-water mixtures, Biochim. Biophys. Acta 38, 64–79, 1980; Zaeklj, A. and Gros, M., Electrophoresis of lipoprotein, prestained with Sudan Black B, dissolved in a mixture of dioxane and ethylene glycol, Clin. Chim. Acta 5, 947, 1960; Krasner, J. and McMenamy, R.H., The binding of indole compounds to bovine plasma albumin. Effects of potassium chloride, urea, dioxane, and glycine, J. Biol. Chem. 241, 4186–4196, 1966; Smith, R.R. and Canady, W.J., Solvation effects upon the thermodynamic substrate activity: correlation with the kinetics of enzyme-catalyzed reactions. II. More complex interactions of alpha-chymotrypsin with dioxane and acetone which are also competitive inhibitors, Biophys. Chem. 43, 189–195, 1992; Forti, F.L., Goissis, G., and Plepis, A.M., Modifications on collagen structures promoted by 1,4-dioxane improve thermal and biological properties of bovine pericardium as a biomaterial, J. Biomater. Appl. 20, 267–285, 2006.


Cleland, W.W., Dithiothreitol, a new protective reagent for SH groups, Biochemistry 3, 480–482, 1964; Gorin, G., Fulford, R., and Deonier, R.C., Reaction of lysozyme with dithiothreitol and other mercaptans, Experientia 24, 26–27, 1968; Stanton, M. and Viswantha, T., Reduction of chymotryptin A by dithiothreitol, Can. J. Biochem. 49, 1233–1235, 1971; Warren, W.A., Activation of serum creatine kinase by dithiothreitol, Clin. Chem. 18, 473–475, 1972; Hase, S. and Walter, R., Symmetrical disulfide bonds as S-protecting groups and their cleavage by dithiothreitol: synthesis of oxytocin with high biological activity, Int. J. Pept. Protein Res. 5, 283–288, 1973; Fleisch, J.H., Krzan, M.C., and Titus, E., Alterations in pharmacologic receptor activity by dithiothreitol, Am. J. Physiol. 227, 1243–1248, 1974; Olsen, J. and Davis, L., The oxidation of dithiothreitol by peroxidases and oxygen, Biochim. Biophys. Acta. 445, 324–329, 1976; Chao, L.P., Spectrophotometric determination of choline acetyltransferase in the presence of dithiothreitol, Anal. Biochem. 85, 20–24, 1978; Fukada, H. and Takahashi, K., Calorimetric study of the oxidation of dithiothreitol, J. Biochem. 87, 1105–1110, 1980; Alliegro, M.C., Effects of dithiothreitol on protein activity unrelated to thiol-disulfide exchange: for consideration in the analysis of protein function with Cleland’s reagent, Anal. Biochem. 282, 102–106, 2000; Rhee, S.S. and Burke, D.H., Tris(2-carboxyethyl)phosphine stabilization of RNA: comparison with dithiothreitol for use with nucleic acid and thiophosphoryl chemistry, Anal. Biochem. 325, 137–143, 2004; Pan, J.C., Cheng, Y., Hui, E.F., and Zhou, H.M., Implications of the role of reactive cysteine in arginine kinase: reactivation kinetics of 5,5′-dithiobis-(2-nitrobenzoic acid)-modified arginine kinase reactivated by dithiothreitol, Biochem. Biophys. Res. Commun. 317, 539–544, 2004; Thaxton, C.S., Hill, H.D., Georganopoulou, D.G. et al., A bio-barcode assay based upon dithiothreitol-induced oligonucleotide release, Anal. Chem. 77, 8174–8178, 2005.


Huggins, C.E., Reversible agglomeration used to remove dimethylsulfoxide from large volumes of frozen blood, Science 139, 504–505, 1963; Yehle, A.V. and Doe, R.H., Stabilization of Bacillus subtilis phage with dimethylsulfoxide, Can. J. Microbiol. 11, 745–746, 1965; Fowler, A.V. and Zabin, I., Effects of dimethylsulfoxide on the lactose operon of Escherichia coli, J. Bacteriol. 92, 353–357, 1966; Williams, A.E. and Vinograd, J., The buoyant behavior of RNA and DNA in cesium sulfate solutions containing dimethylsulfoxide, Biochim. Biophys. Acta 228, 423–439, 1971; Levine, W.G., The effect of dimethylsulfoxide on the binding of 3-methylcholanthrene to rat liver fractions, Res. Commum. Chem. Pathol. Pharmacol. 4, 511–518, 1972; Fink, A.L, The trypsin-catalyzed hydrolysis of N-alpha-benzoyl-L-lysine p-nitrophenyl ester in dimethylsulfoxide at subzero temperatures, J. Biol. Chem. 249, 5072–5932, 1974; Hutton, J.R. and Wetmur, J.G., Activity of endonuclease S1 in denaturing solvents: dimethylsulfoxide, dimethylformamide, formamide, and formaldehyde, Biochem. Biophys. Res. Commun. 66, 942–948, 1975; Gal, A., De Groot, N., and Hochberg, A.A., The effect of dimethylsulfoxide on ribosomal fractions from rat liver, FEBS Lett. 94, 25–27, 1978; Barnett, R.E., The effects of dimethylsulfoxide and glycerol on Na+, K+-ATPase, and membrane structure, Cryobiology 15, 227–229, 1978; Borzini, P., Assali, G., Riva, M.R. et al., Platelet cryopreservation using dimethylsulfoxide/ polyethylene glycol/sugar mixture as cryopreserving solution, Vox Sang. 64, 248–249, 1993; West, R.T., Garza, L.A., II, Winchester, W.R., and Walmsley, J.A., Conformation, hydrogen bonding, and aggregate formation of guanosine 5′-monophosphate and guanosine in dimethylsulfoxide, Nucleic Acids Res. 22, 5128–5134, 1994; Bhattacharjya, S. and Balarma, P., Effects of organic solvents on protein structures; observation of a structured helical core in hen egg-white lysozyme in aqueous dimethylsulfoxide, Proteins 29, 492–507, 1997; Simala-Grant, J.L. and Weiner, J.H., Modulation of the substrate specificity of Escherichia coli dimethylsulfoxide reductase, Eur. J. Biochem. 251, 510–515, 1998; Tsuzuki, W., Ue, A., and Kitamura, Y., Effect of dimethylsulfoxide on hydrolysis of lipase, Biosci. Biotechnol. Biochem. 65, 2078–2082, 2001; Pedersen, N.R., Halling, P.J., Pedersen, L.H. et al., Efficient transesterification of sucrose catalyzed by the metalloprotease thermolysin in dimethylsulfoxide, FEBS Lett. 519, 181–184, 2002; Fan, C., Lu, J., Zhang, W., and Li, G., Enhanced electron-transfer reactivity of cytochrome b5 by dimethylsulfoxide and N,N′-dimethylformamide, Anal. Sci. 18, 1031–1033, 2002; Tait, M.A. and Hik, D.S., Is dimethylsulfoxide a reliable solvent for extracting chlorophyll under field conditions? Photosynth. Res. 78, 87–91, 2003; Malinin, G.I. and Malinin, T.I., Effects of dimethylsulfoxide on the ultrastructure of fixed cells, Biotech. Histochem. 79, 65–69, 2004; Clapisson, G., Salinas, C., Malacher, P. et al., Cryopreservation with hydroxyethylstarch (HES) + dimethylsulfoxide (DMSO) gives better results than DMSO alone, Bull. Cancer 91, E97–E102, 2004.


Lin, T.Y. and Koshland, D.E., Jr., Carboxyl group modification and the activity of lysozyme, J. Biol. Chem. 244, 505–508, 1969; Carraway, K.L., Spoerl, P., and Koshland, D.E., Jr., Carboxyl group modification in chymotrypsin and chymotrypsinogen, J. Mol. Biol. 42, 133–137, 1969; Yamada, H., Imoto, T., Fujita, K. et al., Selective modification of aspartic acid-101 in lysozyme by carbodiimide reaction, Biochemistry 20, 4836–4842, 1981; Buisson, M. and Reboud, A.M., Carbodiimide-induced protein-RNA crosslinking in mammalian subunits, FEBS Lett. 148, 247–250, 1982; Millett, F., Darley-Usmar, V., and Capaldi, R.A., Cytochrome c is crosslinked to subunit II of cytochrome c oxidase by a water-soluble carbodiimide, Biochemistry 21, 3857–3862, 1982; Chen, S.C., Fluorometric determination of carbodiimides with trans-aconitic acid, Anal. Biochem. 132, 272–275, 1983; Davis, L.E., Roth, S.A., and Anderson, B., Antisera specificities to 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide adducts of proteins, Immunology 53, 435–441, 1984; Ueda, T., Yamada, H., and Imoto, T., Highly controlled carbodiimide reaction for the modification of lysozyme. Modification of Leu129 or As119, Protein Eng. 1, 189–193, 1987; Ghosh, M.K., Kildsig, D.O., and Mitra, A.K., Preparation and characterization of methotrexate-immunoglobulin conjugates, Drug. Des. Deliv. 4, 13–25, 1989; Grabarek, Z. and Gergely, J., Zero-length crosslinking procedure with the use of active esters, Anal. Biochem. 185, 131–135, 1990; Gilles, M.A., Hudson, A.Q., and Borders, C.L., Jr., Stability of water-soluble carbodiimides in aqueous solutions, Anal. Biochem. 184, 244–248, 1990; Soinila, S., Mpitsos, G.J., and Soinila, J., Immunohistochemistry of enkephalins: model studies on hapten-carrier conjugates and fixation methods, J. Histochem. Cytochem. 40, 231–239, 1992; Soper, S.A., Hashimoto, M., Situma, C. et al., Fabrication of DNA microarrays onto polymer substrates using UV modification protocols with integration into microfluidic platforms for the sensing of low-abundant DNA point mutations, Methods 37, 103–113, 2005.


Flaschka, H.A., EDTA Titrations: An Introduction to Theory and Practice, Pergammon Press, Oxford, UK, 1964; West, T.S., Complexometry with EDTA and Related Reagents, BDH Chemicals Ltd., Poole (Dorset), UK, 1969; Pribil, R., Analytical Applications of EDTA and Related Compounds, Pergammon Press, Oxford, UK, 1972; Papavassiliou, A.G., Chemical nucleases as probes for studying DNA–protein interactions, Biochem. J. 305, 345–357, 1995; Martell, A.E., and Hancock, R.D., Metal Complexes in Aqueous Solutions, Plenum Press, New York, 1996; Loizos, N. and Darst, S.A, Mapping protein–ligand interactions by footprinting, a radical idea, Structure 6, 691–695, 1998; Franklin, S.J., Lanthanide-mediated DNA hydrolysis, Curr. Opin. Chem. Biol. 5, 201–208, 2001; Heyduk, T., Baichoo, N., and Henduk, E., Hydroxyl radical footprinting of proteins using metal ion complexes, Met. Ions Biol. Syst. 38, 255–287, 2001; Orlikowsky, T.W., Neunhoeffer, F., Goelz, R. et al., Evaluation of IL-8-concentrations in plasma and lyszed EDTA-blood in healthy neonates and those with suspected early onset bacterial infection, Pediatr. Res. 56, 804–809, 2004; Matt, T., Martinez-Yamout, M.A., Dyson, H.J., and Wright, P.E., The CBP/p300 TAZ1 domain in its native state is not a binding partner of MDM2, Biochem. J. 381, 685–691, 2004; Nyborg, J.K. and Peersen, O.B., That zincing feeling: the effects of EDTA on the behavior of zinc-binding transcriptional regulators, Biochem. J. 381, e3–e4, 2004; Haberz, P., Rodriguez-Castanada, F., Junker, J. et al., Two new chiral EDTA-based metal chelates for weak alignment of proteins in solution, Org. Lett. 8, 1275–1278, 2006.


Ellman, G.L., Tissue sulfydryl groups, Arch. Biochem. Biophys. 82, 70–77, 1959; Boyne, A.F. and Ellman, G.L., A methodology for analysis of tissue sulfydryl components, Anal. Biochem. 46, 639–653, 1972; Brocklehurst, K., Kierstan, M., and Little, G., The reaction of papain with Ellman’s reagent (5,5′-dithiobis-(2-nitrobenzoate), Biochem. J. 128, 811–816, 1972; Weitzman, P.D., A critical reexamination of the reaction of sulfite with DTNB, Anal. Biochem. 64, 628–630, 1975; Hull, H.H., Chang, R., and Kaplan, L.J., On the location of the sulfydryl group in bovine plasma albumin, Biochim. Biophys. Acta 400, 132–136, 1975; Banas, T., Banas, B., and Wolny, M., Kinetic studies of the reactivity of the sulfydryl groups of glyceraldehyde-3-phosphate dehydrogenase, Eur. J. Biochem. 68, 313–319, 1976; der Terrossian, E. and Kassab, R., Preparation and properties of S-cyano derivatives of creatine kinase, Eur. J. Biochem. 70, 623–628, 1976; Riddles, P.W., Blakeley, R.L., and Zerner, B., Ellman’s reagent: 5,5′-dithiobis(2-nitrobenzoic acid) — a reexamination, Anal. Biochem. 94, 75–81, 1979; Luthra, N.P., Dunlap, R.B., and Odom, J.D., Characterization of a new sulfydryl group reagent: 6, 6′- diselenobis-(3-nitrobenzoic acid), a selenium analog of Ellman’s reagent, Anal. Biochem. 117, 94–102, 1981; Di Simplicio, P., Tiezzi, A., Moscatelli, A. et al., The SH-SS exchange reaction between the Ellman’s reagent and protein-containing SH groups as a method for determining conformational states: tubulin, Ital. J. Biochem. 38, 83–90, 1989; Woodward, J., Tate, J., Herrmann, P.C., and Evans, B.R., Comparison of Ellman’s reagent with N-(1-pyrenyl)maleimide for the determination of free sulfydryl groups in reduced cellobiohydrolase I from Trichoderma reesei, J. Biochem. Biophys. Methods 26, 121–129, 1993; Berlich, M., Menge, S., Bruns, I. et al., Coumarins give misleading absorbance with Ellman’s reagent suggestive of thiol conjugates, Analyst 127, 333–336, 2002; Riener, C.K., Kada, G., and Gruber, H.J., Quick measurement of protein sulfydryls of Ellman’s reagents and with 4,4′-dithiopyridine, Anal. Bio. Anal. Chem. 373, 266–276, 2002; Zhu, J., Dhimitruka, I., and Pei, D., 5-(2-aminoethyl)dithio-2-nitrobenzoate as a more base-stable alternative to Ellman’s reagent, Org. Lett. 6, 3809–3812, 2004; Owusu-Apenten, R., Colorimetric analysis of protein sulfydryl groups in milk: applications and processing effects, Crit. Rev. Food Sci. Nutr. 45, 1–23, 2005.


Vance, D.E. and Ridgway, N.D., The methylation of phosophatidylethanolamine, Prog. Lipid Res. 27, 61–79, 1988; Louwagie, M., Rabilloud, T., and Garin, J., Use of ethanolamine for sample stacking in capillary electrophoresis, Electrophoresis 19, 2440–2444, 1998; de Nogales, V., Ruiz, R., Roses. M. et al., Background electrolytes in 50% methanol/water for the determination of acidity constants of basic drugs by capillary zone electrophoresis, J. Chromatog. A 1123, 113–120, 2006.


Sela, I., Fluorescence of nucleic acids with ethidium bromide: an indication of the configurative state of nucleic acids, Biochim. Biophys. Acta 190, 216–219, 1969; Le Pecq, J.B., Use of ethidium bromide for separation and determination of nucleic acids of various conformational forms and measurement of their associated enzymes, Methods Biochem. Anal. 20, 41–86, 1971; Borst, P., Ethidium DNA agarose gel electrophoresis: how it started, IUBMB Life 57, 745–747, 2005.


Dufour, E., Bertrand-Harb, C., and Haertle, T., Reversible effects of medium dielectric constant on structural transformation of beta-lactoglobulin and its retinol binding, Biopolymers 33, 589–598, 1993; Escalera, J.B., Bustamante, P., and Martin, A., Predicting the solubility of drugs in solvent mixtures: multiple solubility maxima and the chameleonic effect, J. Pharm. Pharmcol. 46, 172–176, 1994; Gratzer, P.F., Pereira, C.A., and Lee, J.M., Solvent environment modulates effects of glutaraldehyde crosslinking on tissue-derived biomaterials, J. Biomed. Mater. Res. 31, 533–543, 1996; Sepulveda, M.R. and Mata, A.M., The interaction of ethanol with reconstituted synaptosomal plasma membrane Ca2+, Biochim. Biophys. Acta 1665, 75–80, 2004; Ramos, A.S. and Techert, S., Influence of the water structure on the acetylcholinesterase efficiency, Biophys. J. 89, 1990–2003, 2005; Wehbi, Z., Perez, M.D., and Dalgalarrondo, M., Study of ethanol-induced conformation changes of holo and apo alpha-lactalbumin by spectroscopy anad limited proteolysis, Mol. Nutr. Food Res. 50, 34–43, 2006; Sasahara, K. and Nitta, K., Effect of ethanol on folding of hen egg-white lysozyme under acidic condition, Proteins 63, 127–135, 2006; Perham, M., Liao, J., and Wittung-Stafshede, P., Differential effects of alcohol on conformational switchovers in alpha-helical and beta-sheet protein models, Biochemistry 45, 7740–7749, 2006; Pena, M.A., Reillo, A., Escalera, B., and Bustamante, P., Solubility parameter of drugs for predicting the solubility profile type within a wide polarity range in solvent mixtures, Int. J. Pharm. 321, 155–161, 2006; Jenke, D., Odufu, A., and Poss, M., The effect of solvent polarity on the accumulation of leachables from pharmaceutical product containers, Eur. J. Pharm. Sci. 27, 133–142, 2006.


Tanford, C., Buckley, C.E., III, De, P.K., and Lively, E.P., Effect of ethylene glycol on the conformation of gamma-globulin and beta-lactoglobulin, J. Biol. Chem. 237, 1168–1171, 1962; Kay, C.M. and Brahms, J., The influence of ethylene glycol on the enzymatic adenosine triphosphatase activity and molecular conformation of fibrous muscle proteins, J. Biol. Chem. 238, 2945–2949, 1963; Narayan, K.A., The interaction of ethylene glycol with rat-serum lipoproteins, Biochim. Biophys. Acta 137, 22–30, 1968; Bello, J., The state of the tyrosines of bovine pancreatic ribonuclease in ethylene glycol and glycerol, Biochemistry 8, 4535–4541, 1969; Lowe, C.R. and Mosbach, K., Biospecific affinity chromatography in aqueous-organic cosolvent mixtures. The effect of ethylene glycol on the binding of lactate dehydrogenase to an immobilized-AMP analogue, Eur. J. Biochem. 52, 99–105, 1975; Ghrunyk, B.A. and Matthews, C.R., Role of diffusion in the folding of the alpha subunit of tryptophan synthase from Escherichia coli, Biochemistry 29, 2149–2154, 1990; Silow, M. and Oliveberg, M., High concentrations of viscogens decrease the protein folding rate constant by prematurely collapsing the coil, J. Mol. Biol. 326, 263–271, 2003; Naseem, F. and Khan, R.H., Effect of ethylene glycol and polyethylene glycol on the acid-unfolded state of trypsinogen, J. Protein Chem. 22, 677–682, 2003; Hubalek, Z., Protectants used in the cyropreservation of microorganisms, Cryobiology 46, 205–229, 2003; Menezo, Y.J., Blastocyst freezing, Eur. J. Obstet. Gynecol. Reprod. Biol. 155 (Suppl. 1), S12–S15, 2004; Khodarahmi, R. and Yazdanparast, R., Refolding of chemically denatured alpha-amylase in dilution additive mode, Biochim. Biophys. Acta. 1674, 175–181, 2004; Zheng, M., Li, Z., and Huang, X., Ethylene glycol monolayer protected nanoparticles: synthesis, characterization, and interactions with biological molecules, Langmuir 20, 4226–4235, 2004; Bonincontro, A., Cinelli, S., Onori, G., and Stravato, A., Dielectric behavior of lysozyme and ferricytochrome-c in water/ethylene-glycol solutions, Biophys. J. 86, 1118–1123, 2004; Kozer, N. and Schreiber, G., Effect of crowding on protein–protein association rates: fundamental differences between low and high mass crowding agents, J. Mol. Biol. 336, 763–774, 2004; Levin, I., Meiri, G., Peretz, M. et al., The ternary complex of Pseudomonas aeruginosa dehydrogenase with NADH and ethylene glycol, Protein Sci. 13, 1547–1556, 2004; Stupishina, E.A., Khamidullin, R.N., Vylegzhanina, N.N. et al., Ethylene glycol and the thermostability of trypsin in a reverse micelle system, Biochemistry 71, 533–537, 2006; Nordstrom, L.J., Clark, C.A., Andersen, B. et al., Effect of ethylene glycol, urea, and N-methylated glycines on DNA thermal stability: the role of DNA base pair composition and hydration, Biochemistry 45, 9604–9614, 2006.


Raftery, M.A. and Cole, R.D., On the aminoethylation of proteins, J. Biol. Chem. 241, 3457–3461, 1966; Fishbein, L., Detection and thin-layer chromatography of derivatives of ethyleneimine. I. N-carbamoyl and aziridines, J. Chromatog. 26, 522–526, 1967; Yamada, H., Imoto, T., and Noshita, S., Modification of catalytic groups in lysozyme with ethyleneimine, Biochemistry 21, 2187–2192, 1982; Okazaki, K., Yamada, H., and Imoto, T., A convenient S-2-aminoethylation of cysteinyl residues in reduced proteins, Anal. Biochem. 149, 516–520, 1985; Hemminki, K., Reactions of ethyleneimine with guanosine and deoxyguanosine, Chem. Biol. Interact. 48, 249–260, 1984; Whitney, P.L., Powell, J.T., and Sanford, G.L., Oxidation and chemical modification of lung beta-galactosidase-specific lectin, Biochem. J. 238, 683–689, 1986; Simpson, D.M., Elliston, J.F., and Katzenellenbogen, J.A., Desmethylnafoxidine aziridine: an electrophilic affinity label for the estrogen receptor with high efficiency and selectivity, J. Steroid Biochem. 28, 233–245, 1987; Musser, S.M., Pan, S.S., Egorin, M.J. et al., Alkylation of DNA with aziridine produced during the hydrolysis of N,N,N′′-triethylenethiophosphoramide, Chem. Res. Toxicol. 5, 95–99, 1992; Thorwirth, S., Muller, H.S., and Winnewisser, G., The millimeter- and submillimeter-wave spectrum and the dipole moment of ethyleneimine, J. Mol. Spectroso. 199, 116–123, 2000; Burrage, T., Kramer, E., and Brown, F., Inactivation of viruses by azirdines, Dev. Biol. (Basel) 102, 131–139, 2000; Brown, F., Inactivation of viruses by aziridines, Vaccine 20, 322–327, 2001; Sasaki, S., Active oligonucleotides incorporating alkylating agent as potential sequence- and base-selective modifier of gene expression, Eur. J. Pharm. Sci. 13, 43–51, 2001; Hou, X.L., Fan, R.H., and Dai, L.X., Tributylphosphine: a remarkable promoting reagent for the ring-opening reaction of aziridines, J. Org. Chem. 67, 5295–5300, 2002; Thevis, M., Loo, R.R.O., and Loo, J.A., In-gel derivatization of proteins for cysteine-specific cleavages and their analysis by mass spectrometry, J. Proteome Res. 2, 163–172, 2003; Sasaki, M., Dalili, S., and Yudin, A.K., N-arylation of aziridines, J. Org. Chem. 68, 2045–2047, 2003; Gao, G.Y., Harden, J.D., and Zhang, J.P., Cobalt-catalyzed efficient aziridination of alkenes, Org. Lett. 7, 3191–3193, 2005; Hopkins, C.E., Hernandez, G., Lee, J.P., and Tolan, D.R., Aminoethylation in model peptides reveals conditions for maximizing thiol specificity, Arch. Biochem. Biophys. 443, 1–10, 2005; Li, C. and Gershon, P.D., pK(a) of the mRNA cap-specific 2′-O-methyltransferase catalytic lysine by HSQC NMR detection of a two-carbon probe, Biochemistry 45, 907–917, 2006; Vicik R., Helten, H., Schirmeister, T., and Engels, B., Rational design of aziridine-containing cysteine protease inhibitors with improved potency: studies on inhibition mechanism, ChemMedChem, 1, 1021–1028, 2006.


Windmueller, H.G., Ackerman, C.J., and Engel, R.W., Reaction of ethylene oxide with histidine, methionine, and cysteine, J. Biol. Chem. 234, 895–899, 1959; Starbuck, W.C. and Busch, H., Hydroxyethylation of amino acids in plasma albumin with ethylene oxide, Biochim. Biophys. Acta 78, 594–605, 1963; Guengerich, F.P., Geiger, L.E., Hogy, L.L., and Wright,. P.L., In vitro metabolism of acrylonitrile to 2-cyanoethylene oxide, reaction with glutathione, and irreversible binding to proteins and nucleic acids, Cancer Res. 41, 4925–4933, 1981; Peter, H., Schwarz, M., Mathiasch, B. et al., A note on synthesis and reactivity towards DNA of glycidonitrile, the epoxide of acrylonitrile, Carcinogenesis 4, 235–237, 1983; Grammer, L.C. and Patterson, R., IgE against ethylene oxide-altered human serum albumin (ETO-HAS) as an etiologic agent in allergic reactions of hemodialysis patients, Artif. Organs 11, 97–99, 1987; Bolt, H.M., Peter, H., and Fost, U., Analysis of macromolecular ethylene oxide adducts, Int. Arch. Occup. Environ. Health 60, 141–144, 1988; Young, T.L., Habraken, Y., Ludlum, D.B., and Santella, R.M., Development of monoclonal antibodies recognizing 7-(2-hydroxyethyl) guanine and imidazole ring-opened 7-(2-hydroxyethyl) guanine, Carcinogenesis 11, 1685–1689, 1990; Walker, V.E., Fennell, T.R., Boucheron, J.A. et al., Macromolecular adducts of ethylene oxide: a literature review and a time-course study on the formation of 7-(2-hydroxyethyl)guanine following exposure of rats by inhalation, Mutat. Res. 233, 151–164, 1990; Framer, P.B., Bailey, E., Naylor, S. et al., Identification of endogenous electrophiles by means of mass spectrometric determination of protein and DNA adducts, Environ. Health Perspect. 99, 19–24, 1993; Tornqvist, M. and Kautianinen, A., Adducted proteins for identification of endogenous electrophiles, Environ. Health Perspect. 99, 39–44, 1993; Galaev, I. Yu. and Mattiasson, B., Thermoreactive water-soluble polymers, nonionic surfactants, and hydrogels as reagents in biotechnology, Enzyme Microb. Technol. 15, 354–366, 1993; Segerback, D., DNA alkylation by ethylene oxide and mono-substituted expoxides, IARC Sci. Publ. 125, 37–47, 1994; Phillips, D.H. and Farmer, P.B., Evidence for DNA and protein binding by styrene and styrene oxide, Crit. Rev. Toxicol. 24 (Suppl.), S35–S46, 1994; Marczynski, B., Marek, W., and Baur, X., Ethylene oxide as a major factor in DNA and RNA evolution, Med. Hypotheses 44, 97–100, 1995; Mosely, G.A. and Gillis, J.R., Factors affecting tailing in ethylene oxide sterilization part 1: when tailing is an artifact…and scientific deficiencies in ISO 11135 and EN 550, PDA J. Pharm. Sci. Technol. 58, 81–95, 2004.


Lundblad, R.L., Chemical Reagent for Protein Modification, 3rd ed., CRC Press, Boca Raton, FL, 2004; Bowes, T.J. and Gupta, R.S., Induction of mitochondrial fusion by cysteine-alkylators ethyacrynic acid and N-ethylmaleimide, J. Cell Physiol. 202, 796–804, 2005; Engberts, J.B., Fernandez, E., Garcia-Rio, L., and Leis, J.R., Water in oil microemulsions as reaction media for a Diels–Alder reaction between N-ethylmaleimide and cyclopentadiene, J. Org. Chem. 71, 4111–4117, 2006; Engberts, J.B., Fernandez, E., Garcia-Rio, L., and Leis, J.R, AOT-based microemulsions accelerate the 1,3-cycloaddition of benzonitrile oxide to N-ethylmaleimide, J. Org. Chem. 71, 6118–6123, 2006; de Jong, K. and Kuypers, F.A., Sulphydryl modifications alter scramblase activity in murine sickle cell disease, Br. J. Haematol. 133, 427–432, 2006; Martin, H.G., Henley, J.M., and Meyer, G., Novel putative targets of N-ethylmaleimide sensitive fusion proteins (NSF) and alpha/beta soluble NSF attachment proteins (SNAPs) include the Pak-binding nucleotide exchange factor betaPIX, J. Cell. Biochem., 99, 1203–1215, 2006; Carrasco, M.R., Silva, O., Rawls, K.A. et al., Chemoselective alkylation of N-alkylaminooxy-containing peptides, Org. Lett. 8, 3529–3532, 2006; Pobbati, A.V., Stein, A., and Fasshauer, D., N- to C-terminal SNARE complex assembly promotes rapid membrane fusion, Science 313, 673–676, 2006; Mollinedo, F., Calafat, J., Janssen, H. et al., Combinatorial SNARE complexes modulate the secretion of cytoplasmic granules in human neutrophils, J. Immunol. 177, 2831–2841, 2006.


Chadwick, C.S., McEntegart, M.G., and Nairn, R.C., Fluorescent protein tracers; a simple alternative to fluorescein, Lancet 1(7017), 412–414, 1958; Holter, H. and Holtzer, H., Pinocytotic uptake of fluorescein-labeled proteins by various tissue cells, Exp. Cell Res. 18, 421–423, 1959; Schatz, H., Interpretation of Fundus Fluorescein Angiography, Mosby, St. Louis, MO, 1978; Voss, E.W., Fluorescein Hapten: An Immunological Probe, CRC Press, Boca Raton, FL, 1984; Katz, J.N., Gobetti, J.P., and Shipman, C., Jr., Fluorescein dye evaluation of glove integrity, J. Am. Dent. Assoc. 118, 327–331, 1989; Fan, J., Pope, L.E., Vitols, K.S., and Huennekens, F.M., Visualization of folate transport proteins by covalent labeling with fluorescein methotrexate, Adv. Enzyme Regul. 30, 3–12, 1990; Mauger, T.F. and Elson, C.L., Havener’s Ocular Pharmacology, Mosby, St. Louis, MO, 1994; Isaac, P.G., Protocols for Nucleic Acid Analysis by Nonradioactive Probes, Humana Press, Totowa, NJ, 1994; Cavallerano, A.A., Ophthalmic fluorescein angiography, Optom. Clin. 5, 1–23, 1996; Mills, C.O., Milkiewicz, P., Saraswat, V., and Elias, E., Cholyllysyl fluorescein and related lysyl fluorescein conjugated bile acid analogues, Yale J. Biol. Med. 70, 447–457, 1997; Zhang, J., Malicka, J., Gryczynski, I., and Lakowicz, J.R., Surface-enhanced fluorescence of fluorescein-labeled oligonucleotides capping on silver nanoparticles, J. Phys. Chem. B Condens. Matter Mater. Surf. Interfaces Biophys. 109, 7643–7648, 2005; Goldsmith, C.R., Jaworski, J., Sheng, M., and Lippard, S.J., Selective labeling of extracellular proteins containing polyhistidine sequences by a fluorescein-nitrilotriacetic acid conjugate, J. Am. Chem. Soc. 128, 418–419, 2006; Sato, K. and Anzai, J., Fluorometric determination of sugars using fluorescein-labeled concanavalin A-glycogen conjugates, Anal. Bio. Anal. Chem. 384, 1297–1301, 2006; Maes, V., Hultsch, C., Kohl, S. et al., Fluorescein-labeled stable neurotensin derivatives, J. Pept. Sci. 12, 505–508, 2006.


Feldman, M.Y., Reactions of nucleic acids and nucleoproteins with formaldehyde, Prog. Nucleic Acid Res. Mol. Biol. 13, 1–49, 1973; Russell, A.D. and Hopwood, D., The biological uses and importance of glutaraldehyde, Prog. Med. Chem. 13, 271–301, 1976; Means, G.E., Reductive alkylation of amino groups, Methods Enzymol. 47, 469–478, 1977; Winkelhake, J.L., Effects of chemical modification of antibodies on their clearance for the circulation. Addition of simple aliphatic compounds by reductive alkylation and carbodiimide-promoted amide formation, J. Biol. Chem. 252, 1865–1868, 1977; Yamazaki, Y. and Suzuki, H., A new method of chemical modification of N 6-amino group in adenine nucleotides with formaldehyde and a thiol and its application to preparing immobilized ADP and ATP, Eur. J. Biochem. 92, 197–207, 1978; Geoghegan, K.F., Cabacungan, J.C., Dixon, H.B., and Feeney, R.E., Alternative reducing agents for reductive methylation of amino groups in proteins, Int. J. Pept. Protein Res. 17, 345–352, 1981; Kunkel, G.R., Mehradian, M., and Martinson, H.G., Contact-site crosslinking agents, Mol. Cell. Biochem. 34, 3–13, 1981; Fox, C.H., Johnson, F.B., Whiting, J., and Roller, P.P., Formaldehyde fixation, J. Histochem. Cytochem. 33, 845–853, 1985; Conaway, C.C., Whysner, J., Verna, L.K., and Williams, G.M., Formaldehyde mechanistic data and risk assessment: endogenous protection from DNA adduct formation, Pharmacol. Ther. 71, 29–55, 1996; Masuda, N., Ohnishi, T., Kawamoto, S. et al., Analysis of chemical modifications of RNA from formalin-fixed samples and optimization of molecular biology applications for such samples, Nucleic Acids Res. 27, 4436–4443, 1999; Micard, V., Belamri, R., Morel, M., and Guilbert, S., Properties of chemically and physically treated wheat gluten films, J. Agric. Food Chem. 48, 2948–2953, 2000; Taylor, I.A. and Webb, M., Chemical modification of lysine by reductive methylation. A probe for residues involved in DNA binding, Methods Mol. Biol. 148, 301–314, 2001; Perzyna, A., Marty, C., Facopre, M. et al., Formaldehyde-induced DNA crosslink of indolizino[1,2-b]quinolines derived from the A-D rings of camptothecin, J. Med. Chem. 45, 5809–5812, 2002; Yurimoto, H., Hirai, R., Matsuno, N. et al., HxlR, a member of the DUF24 protein family, is a DNA-binding protein that acts as a positive regulator of the formaldehyde-inducible hx1AB operon in Bacillus subtilis, Mol. Microbiol. 57, 511–519, 2005.


Sarkar, P.B., Decomposition of formic acid by periodate, Nature 168, 122–123, 1951; Hass, P., Reactions of formic acid and its salts, Nature 167, 325, 1951; Smillie, L.B. and Neurath, H., Reversible inactivation of trypsin by anhydrous formic acid, J. Biol. Chem 234, 355–359, 1959; Hynninen, P.H. and Ellfolk, N., Use of the aqueous formic acid-chloroform-dimethylformamide solvent system for the purification of porphyrins and hemins, Acta Chem. Scand. 27, 1795–1806, 1973; Heukeshoven, J. and Dernick, R., Reversed-phase high-performance liquid chromatography of virus proteins and other large hydrophobic proteins in formic acid-containing solvents, J. Chromatog. 252, 241–254, 1982; Tarr, G.E. and Crabb, J.W., Reverse-phase high-performance liquid chromatography of hydrophobic proteins and fragments thereof, Anal. Biochem. 131, 99–107, 1983; Heukeshoven, J. and Dernick, R., Characterization of a solvent system for separation of water-insoluble poliovirus proteins by reversed-phase high-performance liquid chromatography, J. Chromatog. 326, 91–101, 1985; De Caballos, M.L., Taylor, M.D., and Jenner, P., Isocratic reverse-phase HPLC separation and RIA used in the analysis of neuropeptides in brain tissue, Neuropeptides 20, 201–209, 1991; Poll, D.J. and Harding, D.R., Formic acid as a milder alternative to trifluoroacetic acid and phosphoric acid in two-dimensional peptide mapping, J. Chromatog. 469, 231–239, 1989; Klunk W.E. and Pettegrew, J.W., Alzheimer’s beta-amyloid protein is covalently modified when dissolved in formic acid, J. Neurochem. 54, 2050–2056, 1990; Erdjument-Bromage, H., Lui, M., Lacomis, L. et al., Examination of the micro-tip reversed phase liquid chromatographic extraction of peptide pools for mass spectrometric analysis, J. Chromatog. A 826, 167–181, 1998; Duewel, H.S. and Honek, J.F., CNBr/formic acid reactions of methionine- and trifluoromethionine-containing lambda lysozyme: probing chemical and positional reactivity and formylation side reactions of mass spectrometry, J. Protein Chem. 17, 337–350, 1998; Kaiser, R. and Metzka, L., Enhancement of cyanogen bromide cleavage yields for methionyl-serine and methionyl-threonine peptide bonds, Anal. Biochem. 266, 1–8, 1999; Rodriguez, J.C., Wong, L., and Jennings, P.A., The solvent in CNBr cleavage reactions determines the fragmentation efficiency of ketosteroid isomerase fusion proteins used in the production of recombinant peptides, Protein Expr. Purif. 28, 224–231, 2003; Zu, Y., Zhao, C., Li, C., and Zhang, L., A rapid and sensitive LC-MS/MS method for determination of coenzyme Q10 in tobacco (Nicotiana tabacum L.) leaves, J. Sep. Sci. 29, 1607–1612, 2006; Kalovidouris, M., Michalea, S., Robola, N. et al., Ultra-performance liquid chromatography/tandem mass spectrometry method for the determination of lercaidipine in human plasma, Rapid Commun. Mass Spectrom., 20, 2939–2946, 2006; Wang, P.G., Wei, J.S., Kim, G. et al., Validation and application of a high-performance liquid chromatography-tandem mass spectrometric method for simultaneous quantification of lopinavir and ritonavir in human plasma using semi-automated 96-well liquid–liquid chromatography, J. Chromatog. A, 1130, 302–307, 2006.


Hopwood, D., Theoretical and practical aspects of glutaraldehyde fixation, Histochem. J., 4, 267–303, 1972; Hassell, J. and Hand, A.R., Tissue fixation with diimidoesters as an alternative to aldehydes. I. Comparison of crosslinking and ultrastructure obtained with dimethylsubserimidate and glutaraldehyde, J. Histochem. Cytochem. 22, 223–229, 1974; Russell, A.D. and Hopwood, D., The biological uses and importance of glutaraldehyde, Prog. Med. Chem. 13, 271–301, 1976; Woodroof, E.A., Use of glutaraldehyde and formaldehyde to process tissue heart valves, J. Bioeng. 2, 1–9, 1978; Heumann, H.G., Microwave-stimulated glutaraldehyde and osmium tetroxide fixation of plant tissue: ultrastructural preservation in seconds, Histochemistry 97, 341–347, 1992; Abbott, L., The use and effects of glutaraldehyde: a review, Occup. Health 47, 238–239, 1995; Jayakrishnan, A. and Jameela, S.R., Glutaraldehyde as a fixative in bioprosthesis and drug delivery matrices, Biomaterials 17, 471–484, 1996; Tagliaferro, P., Tandler, C.J., Ramos, A.J. et al., Immunofluorescence and glutaraldehyde fixation. A new procedure base on the Schiff-quenching method, J. Neurosci. Methods 77, 191–197, 1997; Cohen, R.J., Beales, M.P., and McNeal, J.E., Prostate secretory granules in normal and neoplastic prostate glands: a diagnostic aid to needle biopsy, Hum. Pathol. 31, 1515–1519, 2000; Chae, H.J., Kim, E.Y., and In, M., Improved immobilization yields by addition of protecting agents in glutaraldehyde-induced immobilization of protease, J. Biosci. Bioeng. 89, 377–379, 2000; Nimni, M.E., Glutaraldehyde fixation revisited, J. Long Term Eff. Med. Implants 11, 151–161, 2001; Fujiwara, K., Tanabe, T., Yabuchi, M. et al., A monoclonal antibody against the glutaraldehyde-conjugated polyamine, putrescine: application to immunocytochemistry, Histochem. Cell Biol. 115, 471–477, 2001; Chao, H.H. and Torchiana, D.F., Bioglue: albumin/glutaraldehyde sealant in cardiac surgergy, J. Card. Surg. 18, 500–503, 2003; Migneault, I., Dartiguenave, C., Bertrand, M.J., and Waldron, K.C., Glutaraldehyde: behavior in aqueous solution, reaction with proteins, and application to enzyme crosslinking, Biotechniques 37, 790–796, 2004; Jearanaikoon, S. and Abraham-Peskir, J.V., An x-ray microscopy perspective on the effect of glutaraldehyde fixation on cells, J. Microsc. 218, 185–192, 2005; Buehler, P.W., Boykins, R.A., Jia, Y. et al., Structural and functional characterization of glutaraldehyde-polymerized bovine hemoglobin and its isolated fractions, Anal. Chem. 77, 3466–3478, 2005; Kim, S.S., Lim, S.H., Cho, S.W. et al., Tissue engineering of heart valves by recellularization of glutaraldehyde-fixed porcine values using bone marrow-derived cells, Exp. Mol. Med. 38, 273–283, 2006.


Arias, I.M. and Jakoby, W.B., Glutathione, Metabolism and Function, Raven Press, New York, 1976; Meister, A., Glutamate, Glutamine, Glutathione, and Related Compounds, Academic Press, Orlando, FL, 1985; Sies, H. and Ketterer, B., Glutathione Conjugation: Mechanisms and Biological Significance, Academic Press, London, UK, 1988; Tsumoto, K., Shinoki, K., Kondo, H. et al., Highly efficient recovery of functional single-chain Fv fragments from inclusion bodies overexpressed in Escherichia coli by controlled introduction of oxidizing reagent — application to a human single-chain Fv fragment, J. Immunol. Methods 219, 119–129, 1998; Jiang, X., Ookubo, Y., Fujii, I. et al., Expression of Fab fragment of catalytic antibody 6D9 in an Escherichia coli in vitro coupled transcription/translation system, FEBS Lett. 514, 290–294, 2002; Sun, X.X., Vinci, C., Makmura, L. et al., Formation of disulfide bond in p53 correlates with inhibition of DNA binding and tetramerization, Antioxid. Redox Signal. 5, 655–665, 2003; Sies, H. and Packer, L., Eds., Glutathione Transferases and Gamma-Glutamyl Transpeptidases, Elsevier, Amsterdam, 2005; Smith, A.D. and Dawson, H., Glutathione is required for efficient production of infectious picornativur virions, Virology, 353, 258–267, 2006.


Sarquis, J.L. and Adams, E.T., Jr., The temperature-dependent self-association of beta-lactoglobulin C in glycine buffers, Arch. Biochem. Biophys. 163, 442–452, 1974; Poduslo, J.F., Glycoprotein molecular-weight estimation using sodium dodecyl suflate-pore gradient electrophoresis: comparison of Tris-glycine and Tris-borate-EDTA buffer systems, Anal. Biochem. 114, 131–139, 1981; Patton, W.F., Chung-Welch, N., Lopez, M.F. et al., Tris-tricine and Tris-borate buffer systems provide better estimates of human mesothelial cell intermediate filament protein molecular weights than the standard Trisglycine system, Anal. Biochem. 197, 25–33, 1991; Trasltas, G. and Ford, C.H., Cell membrane antigen-antibody complex dissociation by the widely used glycine-HC1 method: an unreliable procedure for studying antibody internalization, Immunol. Invest. 22, 1–12, 1993; Nail, S.L., Jiang, S., Chongprasert, S., and Knopp, S.A., Fundamentals of freeze-drying, Pharm. Biotechnol. 14, 281–360, 2002; Pyne, A., Chatterjee, K., and Suryanarayanan, R., Solute crystallization in mannitolglycine systems — implications on protein stabilization in freeze-dried formulations, J. Pharm. Sci. 92, 2272–2283, 2003; Hasui, K., Takatsuka, T., Sakamoto, R. et al., Double immunostaining with glycine treatment, J. Histochem. Cytochem. 51, 1169–1176, 2003; Hachmann, J.P. and Amshey, J.W., Models of protein modification in Tris-glycine and neutral pH Bis-Tris gels during electrophoresis: effect of gel pH, Anal. Biochem. 342, 237–245, 2005.


Nakaya, K., Takenaka, O., Horinishi, H., and Shibata, K., Reactions of glyoxal with nucleic acids. Nucleotides and their component bases, Biochim. Biophys. Acta 161, 23–31, 1968; Canella, M. and Sodini, G., The reaction of horse-liver alcohol dehydrogenase with glyoxal, Eur. J. Biochem. 59, 119–125, 1975; Kai, M., Kojima, E., Okhura, Y., and Iwaski, M., High-performance liquid chromatography of N-terminal tryptophan-containing peptides with precolumn fluorescence derivatization with glyoxal, J. Chromatog. A. 653, 235–250, 1993; Murata-Kamiya, N., Kamiya, H., Kayi, H., and Kasai, H., Glyoxal, a major product of DNA oxidation, induces mutations at G:C sites on a shuttle vector plasmid replicated in mammalian cells, Nucleic Acids Res. 25, 1897–1902, 1997; Leng, F., Graves, D., and Chaires, J.B., Chemical crosslinking of ethidium to DNA by glyoxal, Biochim. Biophys. Acta 1442, 71–81, 1998; Thrornalley, P.J., Langborg, A., and Minhas, H.S., Formation of glyoxal, methylglyoxal, and 3-deoxyglucosone in the glycation of proteins by glucose, Biochem. J. 344, 109–116, 1999; Sady, C., Jiang, C.L., Chellan, P. et al., Maillard reactions by alpha-oxoaldehydes: detection of glyoxal-modified proteins, Biochim. Biophys. Acta 1481, 255–264, 2000; Olsen, R., Molander P., Ovrebo, S. et al., Reaction of glyoxal with 2′-deoxyguanosine, 2′-deoxyadenosine, 2′-deoxycytidine, cytidine, thymidine, and calf thymus DNA: identification of the DNA adducts, Chem. Res. Toxicol. 18, 730–739, 2005; Manini, P., La Pietra, P., Panzella, L. et al., Glyoxal formation by Fenton-induced degradation of carbohydrates and related compounds, Carbohydr. Res. 341, 1828–1833, 2006.


Hill, R.L., Schwartz, H.C., and Smith, E.L., The effect of urea and guanidine hydrochloride on activity and optical rotation of crystalline papain, J. Biol. Chem. 234, 572–576, 1959; Appella, E. and Markert, C.L., Dissociation of lactate dehydrogenase into subunits with guanidine hydrochloride, Biochem. Biophys. Res. Commun. 6, 171–176, 1961; von Hippel, P.H. and Wong, K.-Y., On the conformational stability of globular proteins. The effects of various electrolytes and nonelectolytes on the thermal transition ribonuclease transition, J. Biol. Chem. 240, 3909–3923, 1965; Katz, S., Partial molar volume and conformational changes produced by the denaturation of albumin by guanidine hydrochloride, Biochim. Biophys. Acta 154, 468–477, 1968; Shortle, D., Guanidine hydrochloride denaturation studies of mutant forms of staphylococcal nuclease, J. Cell Biochem. 30, 281–289, 1986; Lippke, J.A., Strzempko, M.N., Rai, F.F. et al., Isolation of intact high-molecular-weight DNA by using guanidine isothiocyanate, Appl. Environ. Microbiol. 53, 2588–2589, 1987; Alberti, S. and Fornaro, M., Higher transfection efficiency of genomic DNA purified with a guanidinium thiocyanate–based procedure, Nucleic Acids Res. 18, 351–353, 1990; Shirley, B.A., Urea and guanidine hydrochloride denaturation curves, Methods Mol. Biol. 40, 177–190, 1995; Cota, E. and Clarke, J., Folding of beta-sandwich proteins: three-state transition of a fibronectin type III module, Protein Sci. 9, 112–120, 2000; Kok, T., Wati, S., Bayly, B. et al., Comparison of six nucleic acid extraction methods for detection of viral DNA or RNA sequences in four different non-serum specimen types, J. Clin. Virol. 16, 59–63, 2000; Salamanca, S., Villegas, V., Vendrell, J. et al., The unfolding pathway of leech carboxypeptidase inhibitor, J. Biol. Chem. 277, 17538–17543, 2002; Bhuyan, A.K., Protein stabilization by urea and guanidine hydrochloride, Biochemistry 41, 13386–13394, 2002; Jankowska, E., Wiczk, W., and Grzonka, Z., Thermal and guanidine hydrochloride-induced denaturation of human cystatin C, Eur. Biophys. J. 33, 454–461, 2004; Fuertes, M.A., Perez, J.M., and Alonso, C., Small amounts of urea and guanidine hydrochloride can be detected by a far-UV spectrophotometric method in dialyzed protein solutions, J. Biochem. Biophys. Methods 59, 209–216, 2004; Berlinck, R.G., Natural guanidine derivatives, Nat. Prod. Rep. 22, 516–550, 2005; Rashid, F., Sharma, S., and Bano, B., Comparison of guanidine hydrochloride (GdnHCl) and urea denaturation on inactivation and unfolding of human placental cystatin (HPC), Biophys. J. 91, 686–693, 2006; Nolan, R.L. and Teller, J.K., Diethylamine extraction of proteins and peptides isolated with a mono-phasic solution of phenol and guanidine isothiocyanate, J. Biochem. Biophys. Methods 68, 127–131, 2006.


Good, N.E., Winget, G.D., Winter, W. et al., Hydrogen ion buffers for biological research, Biochemistry 5, 467–477, 1966; Turner, L.V. and Manchester, K.L., Interference of HEPES with the Lowry method, Science 170, 649, 1970; Chirpich, T.P., The effect of different buffers on terminal deoxynucleotidyl transferase activity, Biochim. Biophys. Acta 518, 535–538, 1978; Tadolini, B., Iron autoxidation in MOPS and HEPES buffers, Free Radic. Res. Commun. 4, 149–160, 1987; Simpson, J.A., Cheeseman, K.H., Smith, S.E., and Dean, R.T., Free-radical generation by copper ions and hydrogen peroxide. Stimulation by HEPES buffer, Biochem. J. 254, 519–523, 1988; Abas, L. and Guppy, M., Acetate: a contaminant in HEPES buffer, Anal. Biochem. 229, 131–140, 1995; Schmidt, K., Pfeiffer, S., and Mayer, B., Reaction of peroxynitrite with HEPES or MOPS results in the formation of nitric oxide donors, Free Radic. Biol. Med. 24, 859–862, 1998; Wiedorn, K.H., Olert, J., Stacy, R.A. et al., HOPE — a new fixing technique enables preservation and extraction of high molecular weight DNA and RNA of >20 kb from paraffin-embedded tissues. HEPES-glutamic acid buffer mediated organic solvent protection effect, Pathol. Res. Pract. 198, 735–740, 2002; Fulop, L., Szigeti, G., Magyar, J. et al., Differences in electrophysiological and contractile properties of mammalian cardiac tissues bathed in bicarbonate — and HEPES-buffered solutions, Acta Physiol. Scand.178, 11–18, 2003; Mash, H.E., Chin, Y.P., Sigg, L. et al., Complexation of copper by zwitterionic amino-sulfonic (Good) buffers, Anal. Chem. 75, 671–677, 2003; Sokolowska, M. and Bal, W., Cu(II) complexation by “non-coordinating” N-2-hydroxyethylpiperazine-N′-ethanesulfonic acid (HEPES buffer), J. Inorg. Biochem. 99, 1653–1660, 2005; Zhao, G. and Chasteen, N.D., Oxidation of Good’s buffers by hydrogen peroxide, Anal. Biochem. 349, 262–267, 2006; Hartman, R.F. and Rose, S.D., Kinetics and mechanism of the addition of nucleophiles to alpha,beta-unsaturated thiol esters, J. Org. Chem. 71, 6342–6350, 2006.




Reducing agent; modification of aldehydes and carbohydrates; hydrazinolysis used for release of carbohydrates from protein; derivatives such as dinitrophenylhydrazine used for analysis of carbonyl groups in oxidized proteins; detection of acetyl and formyl groups in proteins.

Schmer, G. and Kreil, G., Micro method for detection of formyl and acetyl groups in proteins, Anal. Biochem. 29, 186–192, 1969; Gershoni, J.M., Bayer, E.A., and Wilchek, M., Blot analyses of glycoconjugates: enzyme-hydrazine — a novel reagent for the detection of aldehydes, Anal. Biochem. 146, 59–63, 1985; O’Neill, R.A., Enzymatic release of oligosaccharides from glycoproteins for chromatographic and electrophoretic analysis, J. Chromatog. A 720, 201–215, 1996; Routier, F.H., Hounsell, E.F., and Rudd, P.M., Quantitation of the oligosaccharides of human serum IgG from patients with rheumatoid arthritis: a critical evaluation of different methods, J. Immunol. Methods 213, 113–130, 1998; Robinson, C.E., Keshavarzian, A., Pasco, D.S. et al., Determination of protein carbonyl groups by immunoblotting, Anal. Biochem. 266, 48–57, 1999; Merry, A.H., Neville, D.C., Royle, L. et al., Recovery of intact 2-aminobenzamide-labeled O-glycans released from glycoproteins by hydrazinolysis, Anal. Biochem. 304, 91–99, 2002; Vinograd, E., Lindner, B., and Seltmann, G., Lipopolysaccharides from Serratia maracescens possess one or two 4-amino-4-deoxy-L-arabinopyranose 1-phosphate residues in the lipid A and D-glycero-D-talo-Oct-ulopyranosonic acid in the inner core region, Chemistry 12, 6692–6700, 2006.



Kawaguchi, M. and Syono, K., The excessive production of indole-3-acetic and its significance in studies of the biosynthesis of this regulator of plant growth and development, Plant Cell Physiol. 37, 1043–1048, 1996; Normanly, J. and Bartel, B., Redundancy as a way of life-IAA metabolism, Curr. Opin. Plant Biol. 2, 207–213, 1999; Leyser, O., Auxin signaling: the beginning, the middle, and the end, Curr. Opin. Plant Biol. 4, 382–386, 2001; Ljung, K., Hull, A.K., Kowalczyk, M. et al., Biosynthesis, conjugation, catabolism, and homeostasis of indole-3-acetic acid in Arabidopsis thaliana, Plant Mol. Biol. 49, 249–272, 2002; Kawano, T. Roles of the reactive oxygen species-generating peroxidase reactions in plant defense and growth induction, Plant Cell Rep. 21, 829–837, 2003; Aloni, R., Aloni, E., Langhans, M., and Ullrich, C.I., Role of cytokine and auxin in shaping root architecture: regulating vascular differentiation, laterial root initiation, root apical dominance, and root gravitropism, Ann. Bot. 97, 882–893, 2006.


The amide is neutral and is not susceptible to either positive or negative influence from locally charged groups; iodoacetamide is frequently used to modify sulfydryl groups as part of reduction and carboxymethylation prior to structural analysis. Crestfield, A.M., Moore, S., and Stein, W.H., The preparation and enzymatic hydrolysis of reduced and S-carboxymethylated proteins, J. Biol. Chem. 238, 622–627, 1963; Watts, D.C., Rabin, B.R., and Crook, E.M., The reaction of iodoacetate and iodoacetamide with proteins as determined with a silver/silver iodide electrode, Biochim. Biophys. Acta 48, 380–388, 1961; Inagami, T., The alkylation of the active site of trypsin with iodoacetamide in the presence of alkylguanidines, J. Biol. Chem. 240, PC3453–PC3455, 1965; Fruchter, R.G. and Crestfield, A.M., The specific alkylation by iodoacetamide of histidine-12 in the active site of ribonuclease, J. Biol. Chem. 242, 5807–5812, 1967; Takahashi, K., The structure and function of ribonuclease T. X. Reactions of iodoacetate, iodoacetamide, and related alkylating reagents with ribonuclease T, J. Biochem. 68, 517–527, 1970; Whitney, P.L., Inhibition and modification of human carbonic anhydrase B with bromoacetate and iodoacetate, Eur. J. Biochem. 16, 126–135, 1970; Harada, M. and Irie, M., Alkylation of ribonuclease from Aspirgillus saitoi with iodoacetate and iodoacetamide, J. Biochem. 73, 705–716, 1973; Halasz, P. and Polgar, L., Effect of the immediate microenvironment on the reactivity of the essential SH group of papain, Eur. J. Biochem. 71, 571–575, 1976; Franzen, J.S., Ishman, P., and Feingold, D.S., Half-of-the-sites reactivity of bovine liver uridine diphosphoglucose dehydrogenase toward iodoacetate and iodoacetamide, Biochemistry 15, 5665–5671, 1976; David, M., Rasched, I.R., and Sund, H., Studies of glutamate dehydrogenase. Methionione-169: the preferentially carboxymethylated residue, Eur. J. Biochem. 74, 379–385, 1977; Ohgi, K., Watanabe, H., Emman, K. et al., Alkylation of a ribonuclease from Streptomyces erthreus with iodoacetate and iodoacetamide, J. Biochem. 90, 113–123, 1981; Dahl, K.H. and McKinley-McKee, J.S., Enzymatic catalysis in the affinity labeling of liver alcohol dehydrogenase with haloacids, Eur. J. Biochem. 118, 507–513, 1981; Syvertsen, C. and McKinley-McKee, J.S., Binding of ligands to the catalytic zinc ion in horse liver alcohol dehydrogenase, Arch. Biochem. Biophys. 228, 159–169, 1984; Communi, D. and Erneux, C., Identification of an active site cysteine residue in type Ins(1,4,5)P35-phosphatase by chemical modification and site-directed mutagenesis, Biochem. J. 320, 181–186, 1996; Sarkany, Z., Skern, T., and Polgar, L., Characterization of the active site thiol group of rhinovirus 21 proteinase, FEBS Lett. 481, 289–292, 2000; Lundblad, R.L., Chemical Reagents for Protein Modification, CRC Press, Boca Raton, FL, 2004.


Traut, R.R., Bollen, A., Sun, T.-T. et al., Methyl-4-mercaptobutyrimidate as a cleavable crosslinking reagent and its application to the Escherichia coli 30S ribosome, Biochemistry 12, 3266–3273, 1973; Schram, H.J. and Dulffer, T., The use of 2-iminothiolane as a protein crosslinking reagent, Hoppe Seylers Z. Physiol.Chem. 358, 137–139, 1977; Jue, R., Lambert, J.M., Pierce, L.R., and Traut, R.R., Addition of sulfyhryl groups Escherichia coli ribosomes by protein modification with 2-iminothiolane (methyl 4-mercaptobutyrimidate), Biochemistry 17, 5399–5406, 1978; Lambert, J.M., Jue, R., and Traut, R.R., Disulfide crosslinking of Escherichia coli ribosomal proteins with 2-iminothiolane (methyl 4-mercaptobutyrimidate): evidence that the crosslinked protein pairs are formed in the intact ribosomal subunit, Biochemistry 17, 5406–5416, 1978; Alagon, A.C. and King, T.P., Activation of polysaccharides with 2-iminothiolane and its use, Biochemistry 19, 4341–4345, 1980; Tolan, D.R. and Traut, R.R., Protein topography of the 40 S ribosomal subunit from rabbit reticulocytes shown by crosslinking with 2-iminothiolane, J. Biol. Chem. 256, 10129–10136, 1981; Boileau, G., Butler, P., Hershey, J.W., and Traut, R.R., Direct crosslinks between initiation factors 1, 2, and 3 and ribosomal proteins promoted by 2-iminothiolane, Biochemistry 22, 3162–3170, 1983; Kyriatsoulis, A., Maly, P., Greuer, B. et al., RNA-protein crosslinking in Escherichia coli ribosomal subunits: localization of sites on 16S RNA which are crosslinked to proteins S17 and S21 by treatment with 2-iminothiolane, Nucleic Acids Res. 14, 1171–1186, 1986; Uchiumi, T., Kikuchi, M., and Ogata, K., Crosslinking study on protein neighborhoods at the subunit interface of rat liver ribosomes with 2-iminothiolane, J. Biol. Chem. 261, 9663–9667, 1986; McCall, M.J., Diril, H., and Meares, C.F., Simplified method for conjugating macrocyclic bifunctional chelating agents to antibodies via 2-iminothiolane, Bioconjug. Chem. 1, 222–226, 1990; Tarentino, A.L., Phelan, A.W., and Plummer, T.H., Jr., 2-iminothiolane: a reagent for the introduction of sulphydryl groups into oligosaccharides derived from asaparagine-linked glycans, Glycobiology 3, 279–285, 1993; Singh, R., Kats, L., Blattler, W.A., and Lambert, J.M., Formation of N-substituted 2-iminothiolanes when amino groups in proteins and peptides are modified by 2-iminothiolanes, Anal. Biochem. 236, 114–125, 1996; Hosono, M.N., Hosono, M., Mishra, A.K. et al., Rhenium-188-labeled anti-neural cell adhesion molecule antibodies with 2-iminothiolane modification for targeting small-cell lung cancer, Ann. Nucl. Med. 14, 173–179, 2000; Mokotoff, M., Mocarski, Y.M., Gentsch, B.L. et al., Caution in the use of 2-iminothiolane (Traut’s reagent) as a crosslinking agent for peptides. The formation of N-peptidyl-2-iminothiolanes with bombesin (BN) antagonists (D-trp6-leu13-ψ[CH2NH]-Phe14BN6-14 and D-trpgln-trp-NH2, J. Pept. Res. 57, 383–389, 2001; Kuzuhara, A., Protein structural changes in keratin fibers induced by chemical modification using 2-iminothiolane hydrochloride: a Raman spectroscopic investigation, Biopolymers 79, 173–184, 2005.


Gelb, M.H. and Abeles, R.H., Substituted isatoic anhydrides: selective inactivators of trypsinlike serine proteases, J. Med. Chem. 29, 585–589, 1986; Gravett, P.S., Viljoen, C.C., and Oosthuizen, M.M., Inactivation of arginine esterase E-1 of Bitis gabonica venom by irreversible inhibitors including a water-soluble carbodiimide, a chloromethyl ketone, and isatoic anhydride, Int. J. Biochem. 23, 1101–1110, 1991; Servillo, L., Balestrieri, C., Quagliuolo, L. et al., tRNA fluorescent labeling at 3′ end including an aminoacyl-tRNA-like behavior, Eur. J. Biochem. 213, 583–589, 1993; Churchich, J.E., Fluorescence properties of o-aminobenzoyl-labeled proteins, Anal. Biochem. 213, 229–233, 1993; Brown, A.D. and Powers, J.C., Rates of thrombin acylation and deacylation upon reaction with low molecular weight acylating agents, carbamylating agents, and carbonylating agents, Bioorg. Med. Chem. 3, 1091–1097, 1995; Matos, M.A., Miranda, M.S., Morais, V.M., and Liebman, J.F., Are isatin and isatoic anhydride antiaromatic and aromatic, respectively? A combined experimental and theoretic investigation, Org. Biomol. Chem. 1, 2566–2571, 2003; Matos, M.A., Miranda, M.S., Morais, V.M., and Liebman, J.F., The energetics of isomeric benzoxazine diones: isatoic anhydride revisited, Org. Biomol. Chem. 2, 1647–1650, 2004; Raturi, A., Vascratsis, P.O., Seslija, D. et al., A direct, continuous, sensitive assay for protein disulphide-isomerase based on fluorescence self-quenching, Biochem. J. 391, 351–357, 2005; Zhang, W., Lu, Y., and Nagashima, T., Plate-to-plate fluorous solid-phase extraction for solution-phase parallel synthesis, J. Comb. Chem. 7, 893–897, 2005.


Brosious, E.M., Morrison, B.Y., and Schmidt, R.M., Effects of hemoglobin F levels, KCN, and storage on the isopropanol precipitation test for unstable hemoglobins, Am. J. Clin. Pathol. 66, 878–882, 1976; Bensinger, T.A. and Beutler, E., Instability of the oxy form of sickle hemoglobin and of methemoglobin in isopropanol, Am. J. Clin. Pathol. 67, 180–183, 1977; Acree, W.E., Jr. and Bertrand, G.L., A cholesterol-isopropanol gel, Nature 269, 450, 1977; Naoum, P.C. Teixeira, U.A., de Abreu Machado, P.E., and Michelin, O.C., The denaturation of human oxyhemoglobin A, A2, and S by isopropanol/buffer method, Rev. Bras. Pesqui. Med. Biol. 11, 241–244, 1978; Ali, M.A., Quinlan, A., and Wong, S.C., Identification of hemoglobin E by the isopropanol solubility test, Clin. Biochem. 13, 146–148, 1980; Horer, O.L. and Enache, C., 2-propanol dependent RNA absorbances, Virologie 34, 257–272, 1983; De Vendittis, E., Masullo, M., and Bocchini, V., The elongation factor G carries a catalytic site for GTP hydrolysis, which is revealed by using 2-propanol in the absence of ribosomes, J. Biol. Chem. 261, 4445–4450, 1986; Wang, L., Hirayasu, K., Ishizawa, M., and Kobayashi, Y., Purification of genomic DNA from human whole blood by isopropanol-fractionation with concentrated NaI and SDS, Nucleic Acids Res. 22, 1774–1775, 1994; Dalhus, B. and Gorbitz, C.H., Glycyl-L-leucyl-L-tyrosine dehydrate 2-propanol solvate, Acta Crystallogr. C 52, 2087–2090, 1996; Freitas, S.S., Santos, J.A., and Prazeres, D.M., Optimization of isopropanol and ammonium sulfate precipitation steps in the purification of plasmid DNA, Biotechnol. Prog. 22, 1179–1186, 2006; Halano, B., Kubo, D., and Tagaya, H., Study on the reactivity of diarylmethane derivatives in supercritical alcohols media: reduction of diarylmethanols and diaryl ketones to diarylmethanes using supercritical 2-propanol, Chem. Pharm. Bull. 54, 1304–1307, 2006.


Cho, S., Scharpf, S., Franko, M., and Vermeulen, C.W., Effect of isopropyl-β-D-galactoside concentration on the level of lac-operon induction in steady state Escherichia coli, Biochem. Biophys. Res. Commun. 128, 1268–1273, 1985; Carlsson, U., Ferskgard, P.O., and Svensson, S.C., A simple and efficient synthesis of the induced IPTG made for inexpensive heterologous protein production using the lac-promoter, Protein Eng. 4, 1019–1020, 1991; Donovan, R.S., Robinson, C.W., and Glick, B.R., Review: optimizing inducer and culture conditions for expression of foreign proteins under control of the lac promoter, J. Ind. Microbiol. 16, 145–154, 1996; Hansen, L.H., Knudsen, S., and Sorensen, S.J., The effect of the lacy gene on the induction of IPTG-inducible promoters, studied in Escherichia coli and Pseudomonas fluorescens, Curr. Microbiol. 36, 341–347, 1998; Teich, A., Lin, H.Y., Andersson, L. et al., Amplification of ColE1 related plasmids in recombinant cultures of Escherichia coli after IPTG induction, J. Biotechnol. 64, 197–210, 1998; Ren, A. and Schaefer, T.S., Isopropyl-β-D-thiogalactoside (IPTG)-inducible tyrosine phosphorylation of proteins in E. coli, Biotechniques 31, 1254–1258, 2001; Ko, K.S., Kruse, J., and Pohl, N.L., Synthesis of isobutryl-C-galactoside (IBCG) as an isopropylthiogalactoside (IPTG) substitute for increased induction of protein expression, Org. Lett. 5, 1781–1783, 2003; Intasai, N., Arooncharus, P., Kasinrerk, W., and Tayapiwatana, C., Construction of high-density display of CD147 ectodomain on VCSM13 phage via gpVIII: effects of temperature, IPTG, and helper phage infection-period, Protein Expr. Purif. 32, 323–331, 2003; Faulkner, E., Barrett, M., Okor, S. et al., Use of fed-batch cultivation for achieving high cell densities for the pilot-scale production of a recombinant protein (phenylalanine dehydrogenase) in Escherichia coli, Biotechnol. Prog. 22, 889–897, 2006; Gardete, S., de Laencastre, H., and Tomasz, A., A link in transcription between the native pbpG and the acquired mecA gene in a strain of Staphylococcus aureus, Microbiology 152, 2549–2558, 2006; Hewitt, C.J., Onyeaka, H., Lewis, G. et al., A comparison of high cell density fed-batch fermentations involving both induced and noninduced recombinant Escherichia coli under well-mixed small-scale and simulated poorly mixed large-scale conditions, Biotechnol. Bioeng., in press, 2006; Picaud, S., Olsson, M.E., and Brodelius, P.E., Improved conditions for production of recombinant plant sesquiterpene synthases in Escherichia coli, Protein Expr. Purif., in press, 2006.


Giese, R.W. and Vallee, B.L., Metallocenes. A novel class of reagents for protein modification. I. Maleic anhydride-iron tetracarbonyl, J. Am. Chem. Soc. 94, 6199–6200, 1972; Cantrell, M. and Craven, G.R., Chemical inactivation of Escherichia coli 30 S ribosomes with maleic anhydride: identification of the proteins involved in polyuridylic acid binding, J. Mol. Biol. 115, 389–402, 1977; Jordano, J., Montero, F., and Palacian, E., Relaxation of chromatin structure upon removal of histones H2A and H2B, FEBS Lett. 172, 70–74, 1984; Jordano, J., Montero, F., and Palacian, E., Rearrangement of nucleosomal components by modification of histone amino groups. Structural role of lysine residues, Biochemistry 23, 4280–4284, 1984; Palacian, E., Gonzalez, P.J., Pineiro, M., and Hernandez, F., Dicarboxylic acid anhydrides as dissociating agents of protein-containing structures, Mol. Cell. Biochem. 97, 101–111, 1990; Paetzel, M., Strynadka, N.C., Tschantz, W.R. et al., Use of site-directed chemical modification to study an essential lysine in Escherichia coli leader peptidase, J. Biol. Chem. 272, 9994–10003, 1997; Wink, M.R., Buffon, A., Bonan, C.D. et al., Effect of protein-modifying reagents on ecto-apyrase from rat brain, Int. J. Biochem. Cell Biol. 32, 105–113, 2000.


Geren, C.R., Olomon, C.M., Jones, T.T., and Ebner, D.E., 2-mercaptoethanol as a substrate for liver alcohol dehydrogenase, Arch. Biochem. Biophys. 179, 415–419, 1977; Opitz, H.G., Lemke, H, and Hewlett, G., Activation of T-cells by a macrophage or 2-mercaptoethanol-activated serum factor is essential for induction of a primary immune response to heterologous red cells in vitro, Immunol. Rev. 40, 53–77, 1978; Burger, M., An absolute requirement for 2-mercaptoethanol in the in vitro primary immune response in the absence of serum, Immunology 37, 669–671, 1979; Nealon, D.A., Pettit, S.M., and Henderson, A.R., Diluent pH and the stability of the thiol group in monothioglycerol, N-acetyl-L-cysteine, and 2-mercaptoethanol, Clin. Chem. 27, 505–506, 1981; Dahl, K.H. and McKinley-McKee, J.S., Enzymatic catalysis in the affinity labeling of liver alcohol dehydrogenase with haloacids, Eur. J. Biochem. 118, 507–513, 1981; Righetti, P.G., Tudor, G., and Glanazza, E., Effect of 2-mercaptoethanol on pH gradients in isoelectric focusing, J. Biochem. Biophys. Methods 6, 219–227, 1982; Soderberg, L.S. and Yeh, N.H., T-cells and the anti-trinitrophenyl antibody response to fetal calf serum and 2-mercaptoethanol, Proc. Soc. Exp. Biol. Med. 174, 107–113, 1983; Ochs, D., Protein contaminants of sodium dodecyl sulfate-polyacrylamide gels, Anal. Biochem. 135, 470–474, 1983; Schaefer, W.H., Harris, T.M., and Guengerich, F.P., Reaction of the model thiol 2-mercaptoethanol and glutathione with methylvinylmaleimide, a Michael acceptor with extended conjugation, Arch. Biochem. Biophys. 257, 186–193, 1987; Obiri, N. and Pruett, S.B., The role of thiols in lymphocyte responses: effect of 2-mercaptoethanol on interleukin 2 production, Immunobiology 176, 440–449, 1988; Gourgerot-Pocidalo, M.A., Fay, M., Roche, Y., and Chollet-Martin, S., Mechanisms by which oxidative injury inhibits the proliferative response of human lymphocytes to PHA. Effect of the thiol compound 2-mercaptoethanol, Immunology 64, 281–288, 1988; Fong, T.C. and Makinodan, T., Preferential enhancement by 2-mercaptoethanol of IL-2 responsiveness of T blast cells from old over young mice is associated with potentiated protein kinase C translocation, Immunol. Lett. 20, 149–154, 1989; De Graan, P.N., Moritz, A., de Wit, M., and Gispen, W.H., Purification of B-50 by 2-mercaptoethanol extraction from rat brain synaptosomal plasma membranes, Neurochem. Res. 18, 875–881, 1993; Carrithers, S.L. and Hoffman, J.L., Sequential methylation of 2-mercaptoenthanol to the dimethyl sulfonium ion, 2-(dimethylthio)ethanol, in vivo and in vitro, Biochem. Pharmacol. 48, 1017–1024, 1994; Paul-Pretzer, K. and Parness, J., Elimination of keratin contaminant from 2-mercaptoethanol, Anal. Biochem. 289, 98–99, 2001; Adebiyi, A.P., Jin, D.H, Ogawa, T., and Muramoto, K., Acid hydrolysis of protein in a microcapillary tube for the recovery of tryptophan, Biosci. Biotechnol. Biochem. 69, 255–257, 2005; Adams, B., Lowpetch, K., Throndycroft, F. et al., Stereochemistry of reactions of the inhibitor/substrates L- and D-β-chloroalanine with β-mercaptoethanol catalyzed by L-aspartate aminotransferase and D-amino acid amino-transferase, respectively, Org. Biomol. Chem. 3, 3357–3364, 2005; Layeyre, M., Leprince, J., Massonneau, M. et al., Aryldithioethyloxycarbonyl (Ardec): a new family of amine-protecting groups removable under mild reducing conditions and their applications to peptide synthesis, Chemistry 12, 3655–3671, 2006; Okun, I., Malarchuk, S., Dubrovskaya, E. et al., Screening for caspace-3 inhibitors: effect of a reducing agent on the identified hit chemotypes, J. Biomol. Screen. 11, 694–703, 2006; Aminian, M., Sivam, S., Lee, C.W. et al., Expression and purification of a trivalent pertussis toxin-diphtheria toxin-tetanus toxin fusion protein in Escherichia coli, Protein Expr. Purif. 51, 170–178, 2006.


Jung, S.K. and Wilson, G.S., Polymeric mercaptosilane-modified platinum electrodes for elimination of interferants in glucose biosensors, Anal. Chem. 68, 591–596, 1996; Mansur, H.S., Lobato, Z.P., Orefice, R.L. et al., Surface functionalization of porous glass networks: effects on bovine serum albumin and porcine insulin immobilization, Biomacromolecules 1, 479–497, 2000; Kumar, A., Larsson, O., Parodi, D., and Liang, Z., Silanized nucleic acids: a general platform for DNA immobilization, Nucleic Acids Res. 28, E71, 2000; Zhang, F., Kang, E.T., Neoh, K.G. et al., Surface modification of stainless steel by grafting of poly(ethylene glycol) for reduction in protein adsorption, Biomaterials 22, 1541–1548, 2001; Jia, J., Wang, B., Wu, A. et al., A method to construct a third-generation horseradish peroxidase biosensor: self-assembling gold nanoparticles to three-dimensional sol-gel network, Anal. Chem. 74, 2217–2223, 2002; AbdelghaniJacquin, C., Abdelghani, A., Chmel, G. et al., Decorated surfaces by biofunctionalized gold beads: application to cell adhesion studies, Eur. Biophys. J. 31, 102–110, 2002; Ganesan, V. and Walcarius, A., Surfactant templated sulfonic acid functionalized silica microspheres as new efficient ion exchangers and electrode modifiers, Langmuir 20, 3632–3640, 2004; Crudden, C.M., Sateesh, M., and Lewis, R., Mercaptopropyl-modified mesoporous silica: a remarkable support for the preparation of a reusable, heterogeneous palladium catalyst for coupling to reactions, J. Am. Chem. Soc. 127, 10045–10050, 2005; Yang, L., Guihen, E., and Glennon, J.D., Alkylthiol gold nanoparticles in sol-gel-based open tabular capillary electrochromatography, J. Sep. Sci. 28, 757–766, 2005.


Good, N.E., Winget, G.D., Winter, W. et al., Hydrogen ion buffers for biological research, Biochemistry 5, 467–477, 1966; Bugbee, B.G. and Salisbury, F.B., An evaluation of MES (2[N-morpholino]ethanesulfonic acid) and Amberlite 1RC-50 as pH buffers for nutrient growth studies, J. Plant Nutr. 8, 567–583, 1985; Kaushal, V. and Barnes, L.D., Effect of zwitterionic buffers on measurement of small masses of protein with bicinchoninic acid, Anal. Biochem. 157, 291–294, 1986; Grady, J.K., Chasteen, N.D., and Harris, D.C., Radicals from “Good’s” buffers, Anal. Biochem. 173, 111–115, 1988; Le Hir, M., Impurity in buffer substances mimics the effect of ATP on soluble 5′-nucleotidase, Enzyme 45, 194–199, 1991; Pedrotti, B., Soffientini, A., and Islam, K., Sulphonate buffers affect the recovery of microtubule-associated proteins MAP1 and MAP2: evidence that MAP1A promotes microtubule assembly, Cell Motil. Cytoskeleton 25, 234–242, 1993; Vasseur, M., Frangne, R., and Alvarado, F., Buffer-dependent pH sensitivity of the fluorescent chloride-indicator dye SPQ, Am. J. Physiol. 264, C27–C31, 1993; Frick, J. and Mitchell, C.A., Stabilization of pH in solid-matrix hydroponic systems, HortScience 28, 981–984, 1993; Yu, Q., Kandegedara, A., Xu, Y., and Rorabacher, D.B., Avoiding interferences from Good’s buffers: a continguous series of noncomplexing tertiary amine buffers covering the entire range of pH 3–11, Anal. Biochem. 253, 50–56, 1997; Gelfi, C., Vigano, A., Curcio, M. et al., Single-strand conformation polymorphism analysis by capillary zone electrophoresis in neutral pH buffer, Electrophoresis 21, 785–791, 2000; Walsh, M.K., Wang, X., and Weimer, B.C., Optimizing the immobilization of single-stranded DNA onto glass beads, J. Biochem. Biophys. Methods 47, 221–231, 2001; Hosse, M. and Wilkinson, K.J., Determination of electrophoretic mobilities and hydrodynamic radii of three humic substances as a function of pH and ionic strength, Environ. Sci. Technol. 35, 4301–4306, 2001; Mash, H.E., Chin, Y.P., Sigg, L. et al., Complexation of copper by zwitterionic aminosulfonic (good) buffers, Anal. Chem. 75, 671–677, 2003; Ozkara, S., Akgol, S., Canak, Y., and Denizli, A., A novel magnetic adsorbent for immunoglobulin-g purification in a magnetically stabilized fluidized bed, Biotechnol. Prog. 20, 1169–1175, 2004; Hachmann, J.P. and Amshey, J.W., Models of protein modification in Tris-glycine and neutral pH Bis-Tris gels during electrophoresis: effect of pH, Anal. Biochem. 342, 237–345, 2005; Krajewska, B. and Ciurli, S., Jack bean (Canavalia ensiformis) urease. Probing acid-base groups of the active site by pH variation, Plant Physiol. Biochem. 43, 651–658, 2005; Zhao, G. and Chasteen, N.D., Oxidation of Good’s buffers by hydrogen peroxide, Anal. Biochem. 349, 262–267, 2006.


Simpson, R.J., Neuberger, M.R., and Liu, T.Y., Complete amino acid analysis of proteins from a single hydrolyzate, J. Biol. Chem. 251, 1936–1940, 1976; Kubota, M., Hirayama, T., Nagase, O., and Yajima, H., Synthesis of two peptides corresponding to an alpha-endophin and gamma-endorphin by the methanesulfonic acid deprotecting procedures, Chem. Pharm. Bull. 27, 1050–1054, 1979; Yajima, H., Akaji, K., Saito, H. et al., Studies on peptides. LXXXII. Synthesis of [4-Gln]-neurotensin by the methanesulfonic acid deprotecting procedure, Chem. Pharm. Bull. 27, 2238–2242, 1979; Sakuri, J. and Nagahama, M. Tryptophan content of Clostridium perfringens epsilon toxin, Infect. Immun. 47, 260–263, 1985; Malmer, M.F. and Schroeder, L.A., Amino acid analysis by high-performance liquid chromatography with methanesulfonic acid hydrolysis and 9-fluorenylmethyl-chloroformate derivatization, J. Chromatog. 514, 227–239, 1990; Weiss, M., Manneberg, M., Juranville, J.F. et al., Effect of the hydrolysis method on the determination of the amino acid composition of proteins, J. Chromatog. A 795, 263–275, 1998; Okimura, K., Ohki, K., Nagai, S., and Sakura, N., HPLC analysis of fatty acyl-glycine in the aqueous methanesulfonic acid hydrolysates of N-terminally fatty acylated peptides, Biol. Pharm. Bull. 26, 1166–1169, 2003; Wrobel, K., Kannamkumarath, S.S., Wrobel, K., and Caruso, J.A., Hydrolysis of proteins with methanesulfonic acid for improved HPLC-ICP-MS determination of seleno-methionine in yeast and nuts, Anal. BioAnal. Chem. 375, 133–138, 2003.


Teale, F.W., Cleavage of haem-protein link by acid methylethylketone, Biochim. Biophys. Acta 35, 543, 1959; Tran, C.D. and Darwent, J.R., Characterization of tetrapyridylporphyrinatozinc (II) apomyoglobin complexes as a potential photosynthetic model, J. Chem. Soc. Faraday Trans. II, 82, 2315–2322, 1986.


Szabo, G., Kertesz, J.C., and Laki, K., Interaction of methylglyoxal with poly-L-lysine, Biomaterials 1, 27–29, 1980; McLaughlin, J.A., Pethig, R., and Szent-Gyorgyi, A., Spectroscopic studies of the protein-methylglyoxal adduct, Proc. Natl. Acad. Sci. USA 77, 949–951, 1980; Cooper, R.A., Metabolism of methylglyoxal in microorganisms, Annu. Rev. Microbiol. 38, 49–68, 1984; Richard, J.P., Mechanism for the formation of methylglyoxal from triosephosphates, Biochem. Soc. Trans. 21, 549–553, 1993; Riley, M.L. and Harding, J.J., The reaction of methylglyoxal with human and bovine lens proteins, Biochim. Biophys. Acta 1270, 36–43, 1995; Thornalley, P.J., Pharmacology of methylglyoxal: formation, modification of proteins and nucleic acids, and enzymatic detoxification — a role in pathogenesis and antiproliferative chemotherapy, Gen. Pharmacol. 27, 565–573, 1996; Nagaraj, R.H., Shipanova, I.N., and Faust, F.M., Protein crosslinking by the Maillard reaction. Isolation, characterization, and in vivo detection of a lysine–lysine crosslink derived from methylglyoxal, J. Biol. Chem. 271, 19338–19345, 1996; Shipanova, I.N., Glomb, M.A., and Nagaraj, R.H., Protein modification by methylglyoxal: chemical nature and synthetic mechanism of a major fluorescent adduct, Arch. Biochem. Biophys. 344, 29–34, 1997; Uchida, K., Khor, O.T., Oya, T. et al., Protein modification by a Maillard reaction intermediate methylglyoxal. Immunochemical detection of fluorescent 5-methylimidazolone derivatives in vivo, FEBS Lett. 410, 313–318, 1997; Degenhardt, T.P., Thorpe, S.R., and Baynes, J.W., Chemical modification of proteins by methylglyoxal, Cell. Mol. Biol. 44, 1139–1145, 1998; Izaguirre, G., Kikonyogo, A., and Pietruszko, R., Methylglyoxal as substrate and inhibitor of human aldehyde dehydrogenase: comparison of kinetic properties among the three isozymes, Comp. Biochem. Physiol. B Biochem. Mol. Biol. 119, 747–754, 1998; Lederer, M.O. and Klaiber, R.G., Crosslinking of proteins by Maillard processes: characterization and detection of lysine–arginine crosslinks derived from glyoxal and methylglyoxal, Bioorg. Med. Chem. 7, 2499–2507, 1999; Kalapos, M.P., Methylglyoxal in living organisms: chemistry, biochemistry, toxicology, and biological implications, Toxicol. Lett. 110, 145–175, 1999; Thornalley, P.J., Landborg, A., and Minhas, H.S., Formation of glyoxal, methylglyoxal, and 3-deoxyglucose in the glycation of proteins by glucose, Biochem. J. 344, 109–116, 1999; Nagai, R., Araki, T., Hayashi, C.M. et al., Identification of N-epsilon(carboxyethyl)lysine, one of the methylglyoxal-derived AGE structures, in glucose-modified protein: mechanism for protein modification by reactive aldehydes, J. Chromatog. B Analyt. Technol. Biomed. Life Sci.788, 75–84, 2003.


Smith, D.J., Maggio, E.T., and Kenyon, G.L., Simple alkanethiol groups for temporary sulfhydryl groups of enzymes, Biochemistry 14, 766–771, 1975; Nishimura, J.S., Kenyon, G.L., and Smith, D.J., Reversible modification of the sulfhydryl groups of Escherichia coli succinic thiokinase with methanethiolating reagents, 5,5′-dithio-bis(2-nitrobenzoic acid), p-hydroxymercuribenzoate, and ethylmercurithiosalicylate, Arch. Biochem. Biophys. 170, 407–430, 1977; Bloxham, D.P., The chemical reactivity of the histidine-195 residue in lactate dehydrogenase thiomethylated at the cysteine-165 residue, Biochem. J. 193, 93–97, 1981; Gavilanes, F., Peterson, D., and Schirch, L., Methyl methanethiosulfate as an active site probe of serine hydroxymethyltransferase, J. Biol. Chem. 257, 11431–11436, 1982; Daly, T.J., Olson, J.S., and Matthews, K.S., Formation of mixed disulfide adducts as cysteine-281 of the lactose repressor protein affects operator- and inducer-binding parameters, Biochemistry 25, 5468–5474, 1986; Salam, W.H. and Bloxham, D.P., Identification of subsidiary catalytic groups at the active site of β-ketoacyl-CoA thiolase by covalent modification of the protein, Biochim. Biophys. Acta 873, 321–330, 1986; Stancato, L.F., Hutchison, K.A., Chakraborti, P.K. et al., Differential effects of the reversible thiol-reactive agents arsenite and methyl methanethiosulfonate on steroid binding by the glucocorticoid receptor, Biochemistry 32, 3739–3736, 1993; Hou, L.X. and Vollmer, S., The activity of S-thiolated modified creatine kinase is due to the regeneration of free thiol at the active site, Biochim. Biophys. Acta 1205, 83–88, 1994; Jensen, P.E., Shanbhag, V.P., and Stigbrand, T., Methanethiolation of the liberated cysteine residues of human α-2-macroglobulin treated with methylamine generates a derivative with similar functional characteristics as native β-2-macroglobulin, Eur. J. Biochem. 227, 612–616, 1995; Trimboli, A.J., Quinn, G.B., Smith, E.T., and Barber, M.J., Thiol modification and site-directed mutagenesis of the flavin domain of spinach NADH: nitrate reductase, Arch. Biochem. Biophys. 331, 117–126, 1996; Quinn, K.E. and Ehrlich, B.E., Methanethiosulfonate derivatives inhibits current through the rynodine receptor/channel, J. Gen. Physiol. 109, 225–264, 1997; Hashimoto, M., Majima, E., Hatanaka, T. et al., Irreversible extrusion of the first loop facing the matrix of the bovine heart mitochondrial ADP/ATP carrier by labeling the Cys(56) residue with the SH-reagent methyl methanethiosulfonate, J. Biochem. 127, 443–449, 2000; Spelta, V., Jiang, L.H., Bailey, R.J. et al., Interaction between cysteines introduced into each transmembrane domain of the rat P2X2 receptor, Br. J. Pharmacol. 138, 131–136, 2003; Britto, P.J., Knipling, L., McPhie, P., and Wolff, J., Thiol-disulphide interchange in tubulin: kinetics and the effect on polymerization, Biochem. J. 389, 549–558, 2005; Miller, C.M., Szegedi, S.S., and Garrow, T.A., Conformation-dependent inactivation of human betaine-homocysteine S-methyltransferase by hydrogen peroxide in vitro, Biochem. J. 392, 443–448, 2005.


Barry, B.W. and Bennett, S.L., Effect of penetration enhancers on the permeation of mannitol, hydrocortisone, and progesterone through human skin, J. Pharm. Pharmacol. 39, 535–546, 1987; Forest, M. and Fournier, A., BOP reagent for the coupling of pGlu and Boc-His(Tos) in solid phase peptide synthesis, Int. J. Pept. Protein Res. 35, 89–94, 1990; Sasaki, H., Kojima, M., Nakamura, J., and Shibasaki, J., Enhancing effect of combining two pyrrolidone vehicles on transdermal drug delivery, J. Pharm. Pharmacol. 42, 196–199, 1990; Uch, A.S., Hesse, U., and Dressman, J.B., Use of 1-methyl-pyrrolidone as a solubilizing agent for determining the uptake of poorly soluble drugs, Pharm. Res. 16, 968–971, 1999; Zhao, F. Bhanage, B.M., Shirai, M., and Arai, M., Heck reactions of iodobenzene and methyl acrylate with conventional supported palladium catalysts in the presence of organic and/or inorganic bases without ligands, Chemistry 6, 843–848, 2000; Lee, P.J., Langer, R., and Shastri, V.P., Role of n-methyl pyrrolidone in the enhancement of aqueous phase transdermal transport, J. Pharm. Sci. 94, 912–917, 2005; Tae, G., Kornfield, J.A., and Hubbell, J.A., Sustained release of human growth hormone from in situ forming hydrogels using self-assembly of fluoroalkyl-ended poly(ethylene glycol), Biomaterials 26, 5259–5266, 2005; Babu, R.J. and Pandit, J.K., Effect of penetration enhancers on the transdermal delivery of bupranolol through rat skin, Drug Deliv. 12, 165–169, 2005; Luan, X. and Bodmeier, R., In situ forming microparticle system for controlled delivery of leupolide acetate: influence of the formulation and processing parameters, Eur. J. Pharm. Sci. 27, 143–149, 2006; Lee, P.J., Ahmad, N., Langer, R. et al., Evaluation of chemical enhancers in the transdermal delivery of lidocaine, Int. J. Pharm. 308, 33–39, 2006; Ruble, G.R., Giardino, O.X., Fossceco, S.L. et al., J. Am. Assoc. Lab. Anim. Sci. 45, 25–29, 2006.


Good, N.E., Winget, G.D., Winter, W. et al., Hydrogen ion buffers for biological research, Biochemistry 5, 467–477, 1966; Altura, B.M., Altura, B.M., Carella, A., and Altura, B.T., Adverse effects of Tris, HEPES, and MOPS buffers on contractile responses of arterial and venous smooth muscle induced by prostaglandins, Prostaglandins Med. 5, 123–130, 1980; Tadolini, B., Iron autoxidation in MOPS and HEPES buffers, Free Radic. Res. Commun. 4, 149–160, 1987; Tadolini, B. and Sechi, A.M., Iron oxidation in MOPS buffer. Effect of phosphorus-containing compounds, Free Radic. Res. Commun. 4, 161–172, 1987; Tadolini, B., Iron oxidation in MOPS buffer. Effect of EDTA, hydrogen peroxide, and FeCl3, Free Radic. Res. Commun. 4, 172–182, 1987; Ishihara, H. and Welsh, M.J., Block by MOPS reveals a conformation change in the CFTR pore produced by ATP hydrolysis, Am. J. Physiol. 273, C1278–C1289, 1997; Schmidt, K., Pfeiffer, S., and Meyer, B., Reaction of peroxynitrite with HEPES or MOPS results in the formation of nitric oxide donors, Free Radic. Biol. Med. 24, 859–862, 1998; Hodges, G.R. and Ingold, K.U., Superoxide, amine buffers, and tetranitromethane: a novel free radical chain reaction, Free Radic. Res. 33, 547–550, 2000; Corona-Izquierdo, F.P. and Membrillo-Hernandez, J., Biofilm formation in Escherichia coli is affected by 3-(N-morpholino)propane sulfonate (MOPS), Res. Microbiol. 153, 181–185, 2002; Mash, H.E., Chin, Y.P., Sigg, L. et al., Complexation of copper by zwitterionic aminosulfonic (Good) buffers, Anal. Chem. 75, 671–677, 2003; Denizli, A., Alkan, M., Garipcan, B. et al., Novel metal-chelate affinity adsorbent for purification of immunoglobulin-G from human plasma, J. Chromatog. B Analyt. Technol. Biomed. Life Sci. 795, 93–103, 2003; Emir, S., Say, R., Yavuz, H., and Denizli, A., A new metal chelate affinity adsorbent for cytochrome C, Biotechnol. Prog. 20, 223–228, 2004; Cvetkovic, A., Zomerdijk, M., Straathof, A.J. et al., Adsorption of fluorescein by protein crystals, Biotechnol. Bioeng. 87, 658–668, 2004; Zhao, G. and Chasteen, J.D., Oxidation of Good’s buffers by hydrogen peroxide, Anal. Biochem. 349, 262–267, 2006; Vrakas, D., Giaginis, C., and TsantiliKakoulidou, A., Different retention behavior of structurally diverse basic and neutral drugs in immobilized artificial membrane and reversed-phase high-performance liquid chromatography: comparison with octanol-water partitioning, J. Chromatog. A 1116, 158–164, 2006; de Carmen Candia-Plata, M., Garcia, J., Guzman, R. et al., Isolation of human serum immunoglobulins with a new salt-promoted adsorbent, J. Chromatog. A 1118, 211–217, 2006.


Sinn, H.J., Schrenk, H.H., Friedrich, E.A. et al., Radioiodination of proteins and lipoproteins using N-bromosuccinimide as oxidizing agent, Anal. Biochem. 170, 186–192, 1988; Tanemura, K., Suzuki, T., Nishida, Y. et al., A mild and efficient procedure for α-bromination of ketones using N-bromosuccinimide catalyzed by ammonium acetate, Chem. Commun. 3, 470–471, 2004; Lundblad, R.L., Chemical Reagents for Protein Modification, 3rd ed., CRC Press, Boca Raton, FL, 2004; Edens, G.J., Redox titration of antioxidant mixtures with N-bromosuccinimide as titrant: analysis by nonlinear least-squares with novel weighting function, Anal. Sci. 21, 1349–1354, 2005; Abdel-Wadood, H.M., Mohamed, H.A., and Mohamed, F.A., Spectrofluorometric determination of acetaminophen with N-bromosuccinimide, J. AOAC Int. 88, 1626–1630, 2005; Krebs, A., Starczyewska, B., Purzanowska-Tarasiewicz, H., and Sledz, J., Spectrophotometric determination of olanzapine by its oxidation with N-bromosuccinimide and cerium(IV) sulfate, Anal. Sci. 22, 829–833, 2006; Braddock, D.C., Cansell, G., Hermitage, S.A., and White, A.J., Bromoiodinanes with a I(III)-Br bond: preparation, X-ray crystallography, and reactivity as electrophilic brominating agents, Chem. Commun. 13, 1442–1444, 2006; Chen, G., Sasaki, M., Li, X., and Yudin, A.K., Strained enamines as versatile intermediates for stereocontrolled construction of nitrogen heterocycles, J. Org. Chem. 71, 6067–6073, 2006; Braddock D.C., Cansell, G., and Hermitage, S.A., Ortho-substituted iodobenzenes as novel organocatalysts for the transfer of electrophilic bromine from N-bromosuccinmide to alkenes, Chem. Commun. 23, 2483–2485, 2006.


Sawicki, W., Kieler, J., and Briand, P., Vital staining with neutral red and trypan blue of 3H-thymidine-labeled cells prior to autoradiography, Stain Technol. 42, 143–146, 1967; Barbosa, P. and Peters, T.M., The effects of vital dyes on living organisms with special reference to methylene blue and neutral red, Histochem. J. 3, 71–93, 1971; Modha, K., Whiteside, J.P., and Spier, R.E., The determination of cellular viability of hybridoma cells in microtitre plates: a colorimetric assay based on neutral red, Cytotechnology 13, 227–232, 1993; Lowik, C.W., Alblas, M.J., van de Ruit, M. et al., Quantification of adherent and nonadherent cell cultured I 96-well plates using the supravital stain neutral red, Anal. Biochem. 213, 426–433, 1993; Ciapetti, G., Granchi, D., Verri, E. et al., Application of a combination of neutral red and amido black staining for rapid, reliable cytotoxicity testing of biomaterials, Biomaterials 17, 1259–1264, 1996; Hall, J.O., Novakofski, J.E., and Beasley, V.R., Neutral red assay modification to prevent cytotoxicity and improve reproducibility using E-63 rat skeletal muscle cells, Biotech. Histochem. 73, 211–221, 1998; Valentin, I., Philippe, M., Lhuguenot, J., and Chagnon, M., Uridine uptake inhibition as a cytotoxicity test for a human hepatoma cell line (HepG2 cells): comparison with the neutral red assay, Toxicology 158, 127–139, 2001; Zuang, V., The neutral red release assay: a review, Altern. Lab. Anim. 29, 575–599, 2001; Choi, J.K. and Yoo, G.S., Fast protein staining in sodium dodecyl sulfate polyacrylamide gel using counter ion-dyes, Coomassie Brilliant Blue R-250 and neutral red, Arch. Pharm. Res. 25, 704–708, 2002; Wang, Z., Zhang, Z., Liu, D., and Dong, S., A temperature-dependent interaction of neutral red with calf thymus DNA, Spectrochim. Acta A Mol. Biomol. Spectrosc. 59, 949–956, 2003; Svendsen, C., Spurgeon, D.J., Hankard, P.K., and Weeks, J.M., A review of lysosomal membrane stability measured by neutral red retention: is it a workable earthworm biomarker? Ecotoxicol. Environ. Saf. 57, 20–29, 2004; Dubrovsky, J.G., Guttenberger, M., Saralegui, A. et al., Neutral red as a probe for confocal scanning microscopy studies of plant roots, Ann. Bot. 97, 1127–1138, 2006; Ni, Y., Lin, D., and Kokot, S., Synchronous fluorescence, UV-visible spectrophotometric, and voltammetric studies of the competitive interaction of bis(1,10-phenanthroline) copper(II) complex and netural red with DNA, Anal. Biochem. 352, 231–242, 2006.


Anderson, G.W., Callahan, F.M., and Zimmerman, J.E., Synthesis of N-hydroxysuccinimide esters of acyl peptides by the mixed anhydride method, J. Am. Chem. Soc. 89, 178, 1967; Lapidot, Y., Rappoport, S., and Wolman, Y., Use of esters of N-hydroxysuccinimide in the synthesis of N-acylamino acids, J. Lipid Res. 8, 142–145, 1967; Holmquist, B., Blumberg, S., and Vallee, B.L., Superactivation of neutral proteases: acylation with N-hydroxysuccinimide esters, Biochemistry 15, 4675–4680, 1976; ‘t Hoen, P.A., de Kort, F., van Ommen, G.J., and den Dunnen, J.T., Fluorescent labeling of cRNA for microarray applications, Nucleic Acids Res. 31, e20, 2003; Vogel, C.W., Preparation of immunoconjugates using antibody oligosaccharide moieties, Methods Mol. Biol. 283, 87–108, 2004; Cooper, M., Ebner, A., Briggs, M. et al., Cy3B: improving the performance of cyanine dyes, J. Fluoresc. 14, 145–150, 2004; Lundblad, R.L., Chemical Reagents for Protein Modification, 3rd ed., CRC Press, Boca Raton, FL, 2004; Zhang, R., Tang, M., Bowyer, A. et al., A novel pH- and ionic-strength-sensitive carboxy methyl dextran hydrogel, Biomaterials 26, 4677–4683, 2005; Tyan, Y.C., Jong, S.B., Liao, J.D. et al., Proteomic profiling of erythrocyte proteins by proteolytic digestion chip and identification using two-dimensional electrospray ionization tandem mass spectrometry, J. Proteome Res. 4, 748–757, 2005; Lovrinovic, M., Spengler, M., Deutsch, C., and Niemeyer, C.M., Synthesis of covalent DNA-protein conjugates by expressed protein ligation, Mol. Biosyst. 1, 64–69, 2005; Smith, G.P., Kinetics of amine modification of proteins, Bioconjug. Chem. 17, 501–506, 2006; Yang, W.C., Mirzael, H., Liu, X., and Regnier, F.E., Enhancement of amino acid detection and quantitation by electrospray ionization mass spectrometry, Anal. Chem. 78, 4702–4708, 2006; Yu, G., Liang, J., He, Z., and Sun, M., Quantum dot-mediated detection of gamma-aminobutyric acid binding sites on the surface of living pollen protoplasts in tobacco, Chem. Biol. 13, 723–731, 2006; Adden, N., Gamble, L.J., Castner, D.G. et al., Phosphonic acid monolayers for binding of bioactive molecules to titanium surfaces, Langmuir 22, 8197–8204, 2006.


Duliere, W.L., The amino-groups of the proteins of human serum. Action of formaldehyde and ninhydrin, Biochem. J. 30, 770–772, 1936; Schwartz, T.B. and Engel, F.L., A photometric ninhydrin method for the measurement of proteolysis, J. Biol. Chem. 184, 197–202, 1950; Troll, W. and Cannan, R.K., A modified photometric ninhydrin method for the analysis of amino and imino acids, J. Biol. Chem. 200, 803–811, 1953; Moore, S. and Stein, W.H., A modified ninhydrin reagent for the photometric determination of amino acids and related compounds, J. Biol. Chem. 211, 907–913, 1954; Rosen, H., A modified ninhydrin colorimetric analysis for amino acids, Arch. Biochem. Biophys. 67, 10–15, 1957; Meyer, H., The ninhydrin reactions and its analytical applications, Biochem. J. 67, 333–340, 1957; Whitaker, J.R., Ninhydrin assay in the presence of thiol compounds, Nature 189, 662–663, 1961; Grant, D.R., Reagent stability in Rosen’s ninhydrin method for analysis of amino acids, Anal. Biochem. 6, 109–110, 1963; Shapiro, R. and Agarwal, S.C., Reaction of ninhydrin with cytosine derivatives, J. Am. Chem. Soc. 90, 474–478, 1968; Moore, S., Amino acid analysis: aqueous dimethylsulfoxide as solvent for the ninhydrin reaction, J. Biol. Chem. 243, 6281–6283, 1968; McGrath, R., Protein measurement by ninhydrin determination of amino acids released by alkaline hydrolysis, Anal. Biochem. 49, 95–102, 1972; Lamothe, P.J. and McCormick, P.G., Role of hydrindantin in the determination of amino acids using ninhydrin, Anal. Chem. 45, 1906–1911, 1973; Quinn, J.R., Boisvert, J.G., and Wood, I., Semi-automated ninhydrin assay of Kjeldahl nitrogen, Anal. Biochem. 58, 609–614, 1974; Chaplin, M.R., The use of ninhydrin as a reagent for the reversible modification of arginine residues in proteins, Biochem. J. 155, 457–459, 1976; Takahashi, K., Specific modification of arginine residues in proteins with ninhydrin, J. Biochem. 80, 1173–1176, 1976; Yu, P.H. and Davis, B.A., Deuterium isotope effects in the ninhydrin reaction of primary amines, Experientia 38, 299–300, 1982; D’Aniello, A., D’Onofrio, G., Pischetola, M., and Strazzulo, L., Effect of various substances on the colorimetric amino acid–ninhydrin reaction, Anal. Biochem. 144, 610–611, 1985; Macchi, F.D., Shen, F.J., Keck, R.G., and Harris, R.J., Amino acid analysis, using postcolumn ninhydrin detection, in a biotechnology laboratory, Methods Mol. Biol. 159, 9–30, 2000; Moulin, M., Deleu, C., Larher, F.R., and Bouchereau, A., High-performance liquid chromatography determination of pipecolic acid after precolumn derivatization using domestic microwave, Anal. Biochem. 308, 320–327, 2002; Pool, C.T., Boyd, J.G., and Tam, J.P., Ninhydrin as a reversible protecting group of amino-terminal cysteine, J. Pept. Res. 63, 223–234, 2004; Schulz, M.M., Wehner, H.D., Reichert, W., and Graw, M., Ninhydrin-dyed latent fingerprints as a DNA source in a murder case, J. Clin. Forensic Med. 11, 202–204, 2004; Buchberger, W. and Ferdig, M., Improved high-performance liquid chromatographic determination of guanidine compounds by precolumn derivatization with ninhydrin and fluorescence detection, J. Sep. Sci. 27, 1309–1312, 2004; Hansen, D.B., and Joullie, M.M., The development of novel ninhydrin analogues, Chem. Soc. Rev. 34, 408–417, 2005.


Fontana, A. and Scofone, E., Sulfenyl halides as modifying reagents for peptides and proteins, Methods Enzymol. 25B, 482–494, 1972; Sanda, A. and Irie, M., Chemical modification of tryptophan residues in ribonuclease form a Rhizopus sp., J. Biochem. 87, 1079–1087, 1980; De Wolf, M.J., Fridkin, M., Epstein, M., and Kohn, L.D., Structure-function studies of cholera toxin and its A and B protomers. Modification of tryptophan residues, J. Biol. Chem. 256, 5481–5488, 1981; Mollier, P., Chwetzoff, S., Bouet, F. et al., Tryptophan 110, a residue involved in the toxic activity but in the enzymatic activity of notexin, Eur. J. Biochem. 185, 263–270, 1989; Cymes, C.D., Iglesias, M.M., and Wolfenstein-Todel, C., Selective modification of tryptophan-150 in ovine placental lactogen, Comp. Biochem. Physiol. B 106, 743–746, 1993; Kuyama, H., Watanabe, M., Toda, C. et al., An approach to quantitate proteome analysis by labeling tryptophan residues, Rapid Commun. Mass Spectrom. 17, 1642–1650, 2003; Lundblad, R.L., Chemical Reagents for Protein Modification, 3rd ed., CRC Press, Boca Raton, FL, 2004; Matsuo, E., Toda, C., Watanabe, M., et al., Selective detection of 2-nitrobenzensulfenyl-labeled peptides by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry using a novel matrix, Proteomics 6, 2042–2049, 2006; Ou, K., Kesuma, D., Ganesan, K. et al., Quantitative labeling of drug-assisted proteomic alterations by combined 2-nitrobenzenesulfenyl chloride (NBS) isotope labeling and 2DE/MS identification, J. Proteome Res. 5, 2194–2206, 2006.




Popular signal from indicator enzymes such as alkaline phosphatase.


Wieme, R.J., van Sande, M., Karcher, D. et al., A modified technique for direct staining with nitro-blue tetrazolium of lactate dehydrogenase iso-enzyme upon agar gel electrophoresis, Clin. Chim. Acta 7, 750–754, 1962; DeBari, V.A., Coste, J.F., and Needle, M.A., Direct spectrophotometric observation of intracellular nitro-blue tetrazolium and its formazan by multiple internal reflectance infrared spectroscopy, Histochemistry 45, 83–88, 1975; Fried, R., Enzymatic and nonenzymatic assay of superoxide dismutase, Biochimie 57, 657–660, 1975; DeBari, V.A. and Needle, M.A., Mechanism for transport of nitro-blue tetrazolium into viable and nonviable leukocytes, Histochemistry 56, 155–163, 1978; Ellsaesser, C., Miller, N., Lobb, C.J., and Clem, L.W., A new method for the cytochemical staining of cells immobilized in agarose, Histochemistry 80, 559–562, 1984; Walker, S.W., Howie, A.F., and Smith, A.F., The measurement of glycosylated albumin by reduction of alkaline nitro-blue tetrazolium, Clin. Chim. Acta 156, 197–206, 1986; Stegmaier, K., Corsello, S.M., Ross, K.N. et al., Gefitinib induces myeloid differentiation of acute myeloid leukemia, Blood 106, 2841–2848, 2005.


Marland, J.S. and Mulley, B.A., A phase-rule study of multiple-phase formation in a model emulsion system containing water, n-octanol, n-dodecane, and a non-ionic surface-active agent at 10 and 25 degrees, J. Pharm. Pharmacol. 23, 561–572, 1971; Dorsey, J.G. and Khaledi, M.G., Hydrophobicity estimations by reversed-phase liquid chromatography. Implications for biological partitioning processes, J. Chromatog. 656, 485–499, 1993; Vailaya, A. and Horvath, C., Retention in reversed-phase chromato-graphy: partition or adsorption? J. Chromatog. 829, 1–27, 1998; Kellogg, G.E. and Abraham, D.J., Hydrophobicity: is logP(o/w) more than the sum of its parts? Eur. J. Med. Chem. 35, 651–661, 2000; van de Waterbeemd,H., Smith, D.A., and Jones, B.C., Lipophilicity in PK design: methyl, ethyl, futile, J. Comput. Aided Mol. Des. 15, 273–286, 2001; Bethod, A. and Carda-Broch, S., Determination of liquid–liquid partition coefficients by separation methods, J. Chromatog. A 1037, 3–14, 2004.


Hoch, F.L., Willams, R.J., and Vallee, B.L., The role of zinc in alcohol dehydrogenases. II. The kinetics of the instantaneous reversible inactivation of yeast alcohol dehydrogenase by 1,10-phenanthroline, J. Biol. Chem. 232, 453–464, 1958; Sigman, D.S. and Chen, C.H., Chemical nucleases: new reagents in molecular biology, Annu. Rev. Biochem. 59, 207–236, 1990; Pan, C.Q., Landgraf, R., and Sigman, D.S., DNA-binding proteins as site-specific nucleases, Mol. Microbiol. 12, 335–342, 1994; Galis, Z.S., Sukhova, G.K., and Libby, P., Microscopic localization of active proteases by in situ zymography: detection of matrix metalloproteinase activity in vascular tissue, FASEB J. 9, 974–980, 1995; Papavassiliou, A.G., Chemical nucleases as probes for studying DNA–protein interactions, Biochem. J. 305, 345–357, 1995; Perrin, D.M., Mazumder, A., and Sigman, D.S., Oxidative chemical nucleases, Prog. Nucleic Acid Res. Mol. Biol. 52, 123–151, 1996; Sigman, D.S., Landgraf, R., Perrin, D.M., and Pearson, L., Nucleic acid chemistry of the cuprous complexes of 1,10-phenanthroline and derivatives, Met. Ions Biol. Syst. 33, 485–513, 1996; Cha, J., Pedersen, M.V., and Auld, D.S., Metal and pH dependence of heptapeptide catalysis by human matrilysin, Biochemistry 35, 15831–15838, 1996; Kidani, Y. and Hirose, J., Coordination chemical studies on metalloenzymes. II. Kinetic behavior of various types of chelating agents towards bovine carbonic anhydrase, J. Biochem. 81, 1383–1391, 1997; Marini, I., Bucchioni, L., Borella, P. et al., Sorbitol dehydrogenase from bovine lens: purification and properties, Arch. Biochem. Biophys. 340, 383–391, 1997; Dri, P., Gasparini, C., Menegazzi, R. et al., TNF-induced shedding of TNF receptors in human polymorphonuclear leukocytes: role of the 55-kDa TNF receptor and involvement of a membrane-bound and non-matrix metalloproteinase, J. Immunol. 165, 2165–2172, 2000; Kito, M. and Urade, R., Protease activity of 1,10-phenanthroline-copper systems, Met. Ions Biol. Syst. 38, 187–196, 2001; Winberg, J.O., Berg, E., Kolset, S.O. et al., Calcium-induced activation and truncation of promatrix metalloproteinase-9 linked to the core protein of chondroitin sulfate proteoglycans, Eur. J. Biochem. 270, 3996–4007, 2003; Butler, G.S., Tam, E.M., and Overall, C.M., The canonical methionine 392 of matrix metalloproteinase 2 (gelatinase A) is not required for catalytic efficiency or structural integrity: probing the role of the methionine-turn in the metzincin metalloprotease superfamily, J. Biol. Chem. 279, 15615– 15620, 2004; Vauquelin, G. and Vanderheyden, P.M., Metal ion modulation of cystinyl aminopeptidase, Biochem. J. 390, 351–357, 2005; Schilling, S., Cynis, H., von Bohlen, A. et al., Isolation, catalytic properties, and competitive inhibitors of the zinc-dependent murine glutaminyl cyclase, Biochemistry 44, 13415–13424, 2005; Vik, S.B. and Ishmukhametov, R.R., Structure and function of subunit a of the ATP synthase of Escherichia coli, J. Bioenerg. Biomembr. 37, 445–449, 2005.


Braun, W., Burrous, J.W., and Phillips, J.H., Jr., A phenol-extracted bacterial deoxyribonucleic acid, Nature 180, 1356–1357, 1957; Habermann, V., Evidence for peptides in RNA prepared by phenol extraction, Biochim. Biophys. Acta 32, 297–298, 1959; Colter, J.S., Brown, R.A., and Ellem, K.A., Observations on the use of phenol for the isolation of deoxyribonucleic acid, Biochim. Biophys. Acta 55, 31–39, 1962; Lust, J. and Richards, V., Influence of buffers on the phenol extraction of liver microsomal ribonucleic acids, Anal. Biochem. 20, 65–76, 1967; Yamaguchi, M., Dieffenbach, C.W., Connolly, R. et al., Effect of different laboratory techniques for guanidinium-phenol-chloroform RNA extraction on A260/A280 and on accuracy of mRNA quantitation by reverse transcriptase-PCR, PCR Methods Appl. 1, 286–290, 1992; Pitera, R., Pitera, J.E., Mufti, G.J., Salisbury, J.R., and Nickoloff, J.A., Sepharose spin column chromatography. A fast, nontoxic replacement for phenol: chloroform extraction/ethanol precipitation, Mol. Biotechnol. 1, 105–108, 1994; Finnegan, M.T., Herbert, K.E., Evans, M.D., and Lunec, J., Phenol isolation of DNA yields higher levels of 8-deoxodeoxyguanosine compared to pronase E isolation, Biochem. Soc. Trans. 23, 430S, 1995; Beaulieux, F., See, D.M., Leparc-Goffart, I. et al., Use of magnetic beads versus guanidium thiocyanate-phenol-chloroform RNA extraction followed by polymerase chain reaction for the rapid, sensitive detection of enterovirus RNA, Res. Virol. 148, 11–15, 1997; Fanson, B.G., Osmack, P., and Di Bisceglie, A.M., A comparison between the phenol-chloroform method of RNA extraction and the QIAamp viral RNA kit in the extraction of hepatitis C and GB virus-C/hepatitis G viral RNA from serum, J. Virol. Methods 89, 23–27, 2000; Kochl, S., Niederstratter, N., and Parson, W., DNA extraction and quantitation of forensic samples using the phenol-chloroform method and real-time PCR, Methods Mol. Biol. 297, 13–30, 2005; Izzo, V., Notomista, E., Picardi, A. et al., The thermophilic archaeon Sulfolobus solfatarius is able to grow on phenol, Res. Microbiol. 156, 677–689, 2005; Robertson, N. and Leek, R., Isolation of RNA from tumor samples: single-step guanidinium acid-phenol method, Methods Mol. Biol. 120, 55–59, 2006.


Nakahishi, M., Wilson, A.C., and Nolan, R.A., Phenoxyethanol: protein preservative for taxonomists, Science 163, 681–683, 1969; Frolich, K.W., Anderson, L.M., Knutsen, A., and Flood, P.R., Phenoxyethanol as a nontoxic substitute for formaldehyde in long-term preservation of human anatomical specimens for dissection and demonstration purposes, Anat. Rec. 208, 271–278, 1984.


Takahashi, K., The reaction of phenylglyoxal with arginine residues in proteins, J. Biol. Chem. 243, 6171–6179, 1968; Bunzli, H.F. and Bosshard, H.R., Modification of the single arginine residue in insulin with phenylglyoxal, Hoppe Seylers Z. Physiol. Chem. 352, 1180–1182, 1971; Cheung, S.T. and Fonda, M.L., Reaction of phenylglyoxal with arginine. The effect of buffers and pH, Biochem. Biophys. Res. Commun. 90, 940–947, 1979; Srivastava, A. and Modak, M.J., Phenylglyoxal as a template site-specific reagent for DNA or RNA polymerases. Selective inhibition of initiation, J. Biol. Chem. 255, 917–921, 1980; Communi, D., Lecocq, R., Vanweyenberg, V., and Erneux, C., Active site labeling of inositol 1,4,5-triphosphate 3-kinase A by phenylglyoxal, Biochem. J. 310, 109–115, 1995; Eriksson, O., Fontaine, E., and Bernardi, P., Chemical modification of arginines by 2,3-butanedione and phenylglyoxal causes closure of the mitochondrial permeability transition pore, J. Biol. Chem. 273, 12669–12674, 1998; Redowicz, M.J., Phenylglyoxal reveals phosphorylation-dependent difference in the conformation of Acanthamoeba myosin II active site, Arch. Biochem. Biophys. 384, 413–417, 2000; Kucera, I., Inhibition by phenylglyoxal of nitrate transport in Paracoccus denitrificans; a comparison with the effect of a protonophorous uncoupler, Arch. Biochem. Biophys. 409, 327–334, 2003; Johans, M., Milanesi, E., Frank, M. et al., Modification of permeability transition pore arginine(s) by phenylglyoxal derivatives in isolated mitochondria and mammalian cells. Structure-function relationship of arginine ligands, J. Biol. Chem. 280, 12130–12136, 2005.


Wilchek, M., Ariely, S., and Patchornik, A., The reaction of asparagine, glutamine, and derivatives with phosgene, J. Org. Chem. 33, 1258–1259, 1968; Hamilton, R.D. and Lyman, D.J., Preparation of N-carboxy-α-amino acid anhydrides by the reaction of copper(II)-amino acid complexes with phosgene, J. Org. Chem. 34, 243–244, 1969; Pohl, L.R., Bhooshan, B., Whittaker, N.F., and Krishna, G., Phosgene: a metabolite of chloroform, Biochem. Biophys. Res. Commun. 79, 684–691, 1977; Gyllenhaal, O., Derivatization of 2-amino alcohols with phosgene in aqueous media: limitations of the reaction selectivity as found in the presence of O-glucuronides of alprenolol in urine, J. Chromatog. 413, 270–276, 1987; Gyllenhaal, O. and Vessman, J., Phosgene as a derivatizing reagent prior to gas and liquid chromatography, J. Chromatog. 435, 259–269, 1988; Noort, D., Hulst, A.G., Fidder, A., et al. In vitro adduct formation of phosgene with albumin and hemoglobin in human blood, Chem. Res. Toxicol. 13, 719–726, 2000; Lemoucheux, L. Rouden, J., Ibazizene, M. et al., Debenylation of tertiary amies using phosgene or triphosgen: an efficient and rapid procedure for the preparation of carbamoyl chlorides and unsymmetrical ureas. Application in carbon-11 chemistry, J. Org. Chem. 68, 7289–7297, 2003.

Picric Acid



Analytical reagent.

De Wesselow, O.L., The picric acid method for the estimation of sugar in blood and a comparison of this method with that of MacLean, Biochem. J. 13, 148–152, 1919; Newcomb, C., The error due to impure picric acid in creatinine estimations, Biochem. J. 18, 291–293, 1924; Davidsen, O., Fixation of proteins after agarose gel electrophoresis by means of picric acid, Clin. Chim. Acta 21, 205–209, 1968; Gisin, B.F., The monitoring of reactions in solid-phase peptide synthesis with picric acid, Anal. Chim. Acta 58, 248–249, 1972; Hancock, W.S., Battersby, J.E., and Harding, D.R., The use of picric acid as a simple monitoring procedure for automated peptide synthesis, Anal. Biochem. 69, 497–503, 1975; Vasiliades, J., Reaction of alkaline sodium picrate with creatinine: I. Kinetics and mechanism of formation of the mono-creatinine picric acid complex, Clin. Chem. 22, 1664–1671, 1976; Somogyi, P. and Takagi, H., A note on the use of picric acid-formaldehyde-glutaraldehyde fixative for correlated light and electron microscopic immunocytochemistry, Neuroscience 7, 1779–1783, 1982; Meyer, M.H., Meyer, R.A., Jr., Gray, R.W., and Irwin, R.L., Picric acid methods greatly overestimate serum creatinine in mice: more accurate results with high-performance liquid chromatography, Anal. Biochem. 144, 285–290, 1985; Knisley, K.A. and Rodkey, L.S., Direct detection of carrier ampholytes in immobilized pH gradients using picric acid precipitation, Electrophoresis 13, 220–224, 1992; Massoomi, F., Mathews, H.G., III, and Destache, C.J., Effect of seven fluoroquinolines on the determination of serum creatinine by the picric acid and enzymatic methods, Ann. Pharmacother. 27, 586–588, 1993.


Morin, L.G. and Prox, J., New and rapid procedure for serum phosphorus using o-phenylenediamine as reductant, Clin. Chim. Acta. 46, 113–117, 1973; Ohnishi, S.T. and Gall, R.S., Characterization of the catalyzed phosphate assay, Anal. Biochem. 88, 347–356, 1978; Steige, H. and Jones, J.D., Determination of serum inorganic phosphorus using a discrete analyzer, Clin. Chim. Acta. 103, 123–127, 1980, Plaizier-Vercammen, J.A. and De Neve, R.E., Interaction of povidone with aromatic compounds. II: evaluation of ionic strength, buffer concentration, temperature, and pH by factorial analysis, J. Pharm. Sci. 70, 1252–1256, 1981; van Zanten, A.P. and Weber, J.A., Direct kinetic method for the determination of phosphate, J. Clin. Chem. Clin. Biochem. 25, 515–517, 1987; Barlow, I.M., Harrison, S.P., and Hogg, G.L., Evaluation of the Technicon Chem-1, Clin. Chem. 34, 2340–2344, 1988; Giulliano, K.A., Aqueous two-phase protein partitioning using textile dyes as affinity ligands, Anal. Biochem. 197, 333–339, 1991; Goldenheim, P.D., An appraisal of povidone-iodine and wound healing, Postgrad. Med. J., 69 (Suppl. 3), S97–S105, 1993; Vemuri, S., Yu, C.D., and Roosdorp, N., Effect of cryoprotectants on freezing, lyophilization, and storage of lyophilized recombinant alpha 1-antitrypsin formulations, PDA J. Pharm. Sci. Technol. 48, 241–246, 1994; Anchordoquy, T.J. and Carpenter, J.F., Polymers protect lactate dehydrogenase during freeze-drying by inhibiting dissociation in the frozen state, Arch. Biochem. Biophys. 332, 231–238, 1996; Fleisher, W., and Reimer, K., Povidone-iodine in antisepsis — state of the art, Dermatology 195 (Suppl. 2), 3–9, 1997; Fernandes, S., Kim, H.S., and Hatti-Kaul, R., Affinity extraction of dye- and metal ion-binding proteins in polyvinalypyrrolidone-based aqueous two-phase system, Protein Expr. Purif. 24, 460–469, 2002; D’Souza, A.J., Schowen, R.L., Borchardt, R.T. et al., Reaction of a peptide with polyvinylpyrrolidone in the solid state, J. Pharm. Sci. 92, 585–593, 2003; Kaneda, Y., Tsutsumi, Y., Yoshioka, Y. et al., The use of PVP as a polymeric carrier to improve the plasma half-life of drugs, Biomaterials 25, 3259–3266, 2004; Art, G., Combination povidone-iodine and alcohol formulations more effective, more convenient versus formulations containing either iodine or alcohol alone: a review of the literature, J. Infus. Nurs. 28, 314–320, 2005; Yoshioka, S., Aso, Y., and Miyazaki, T., Negligible contribution of molecular mobility to the degradation of insulin lyophilized with poly(vinylpyrrolidone), J. Pharm. Sci. 95, 939–943, 2006.


Klingsberg, E. and Newkome, G.R., Eds., Pyridine and Its Derivatives, Interscience, New York, 1960; Schoefield, K., Hetero-aromatic Nitrogen Compounds; Pyrroles and Pyridines, Butterworths, London, 1967; Hurst, D.T., An Introduction to the Chemistry and Biochemistry and Pyrimidines, Purines, and Ptreridines, J. Wiley, Chichester, UK, 1980; Plunkett, A.O., Pyrrole, pyrrolidine, pyridine, piperidine, and azepine alkaloids, Nat. Prod. Rep. 11, 581–590, 1994; Kaiser, J.P., Feng, Y., and Bollag, J.M., Microbial metabolism of pyridine, quinoline, acridine, and their derivatives under aerobic and anaerobic conditions, Microbiol. Rev. 60, 483–498, 1996.


Hughes, R.C., Jenkins, W.T., and Fischer, E.H., The site of binding of pyridoxal-5′-phosphate to heart glutamic-aspartic transaminase, Proc. Natl. Acad. Sci. USA 48, 1615–1618, 1962; Finseth, R. and Sizer, I.W., Complexes of pyridoxal phosphate with amino acids, peptides, polylysine, and apotransaminase, Biochem. Biophys. Res. Commun. 26, 625–630, 1967; Pages, R.C., Benditt, E.P., and Kirkwood, C.R., Schiff base formation by the lysyl and hydroxylysyl side chains of collagen, Biochem. Biophys. Res. Commun. 33, 752–757, 1968; Whitman, W.B., Martin, M.N., and Tabita, F.R., Activation and regulation of ribulose bisphosphate carboxylase-oxygenase in the absence of small subunits, J. Biol. Chem. 254, 10184–10189, 1979; Howell, E.E. and Schray, K.J., Comparative inactivation and inhibition of the anomerase and isomerase activities of phosphoglucose isomerase, Mol. Cell. Biochem. 37, 101–107, 1981; Colanduoni, J. and Villafranca, J.J., Labeling of specific lysine residues at the active site of glutamine synthetase, J. Biol. Chem. 260, 15042–15050, 1985; Peterson, C.B., Noyes, C.M., Pecon, J.M. et al., Identification of a lysyl residue in antithrombin which is essential for heparin binding, J. Biol. Chem. 262, 8061–8065, 1987; Diffley, J.F., Affinity labeling the DNA polymerase alpha complex. Identification of subunits containing the DNA polymerase active site and an important regulatory nucleotide-binding site, J. Biol. Chem. 263, 19126–19131, 1988; Perez-Ramirez, B. and Martinez-Carrion, M., Pyridoxal phosphate as a probe of the cytoplasmic domains of transmembrane proteins: application to the nicotinic acetylcholine receptor, Biochemistry 28, 5034–5040, 1989; Valinger, Z., Engel, P.C., and Metzler, D.E., Is pyridoxal-5′-phosphate an affinity label for phosphate-binding sites in proteins? The case of bovine glutamate dehydrogenase, Biochem. J. 294, 835–839, 1993; Illy, C., Thielens, N.M., and Arlaud, G.J., Chemical characterization and location of ionic interactions involved in the assembly of the C1 complex of human complement, J. Protein Chem. 12, 771–781, 1993; Hountondji, C., Gillet, S., Schmitter, J.M. et al., Affinity labeling of Escherichia coli lysyl-tRNA synthetase with pyridoxal mono- and diphosphate, J. Biochem. 116, 502–507, 1994; Brody, S., Andersen, J.S., Kannangara, C.G. et al., Characterization of the different spectral forms of glutamate-1-semialdehyde aminotransferase by mass spectrometry, Biochemistry 34, 15918–15924, 1995; Kossekova, G., Miteva, M., and Atanasov, B., Characterization of pyridoxal phosphate as an optical label for measuring electrostatic potentials in proteins, J. Photochem. Photobiol. B 32, 71–79, 1996; Kim S.W., Lee, J., Song, M.S. et al., Essential active-site lysine of brain glutamate dehydrogenase isoproteins, J. Neurochem. 69, 418–422, 1997; Martin, D.L., Liu, H., Martin, S.B., and Wu, S.J., Structural features and regulatory properties of the brain glutamate decarboxylase, Neurochem. Int. 37, 111–119, 2000; Jaffe, M. and Bubis, J., Affinity labeling of the guanine nucleotide binding site of transducin by pyridoxal 5′-phosphate, J. Protein Chem. 21, 339–359, 2002.

Sodium Borohydride



Reducing agent for Schiff bases; reduction of aldehydes; other chemical reductions.

Chaykin, S., King, L., and Watson, J.G., The reduction of DPN+ and TPN+ with sodium borohydride, Biochim. Biophys. Acta 124, 13–25, 1966; Cerutti, P. and Miller, N., Selective reduction of yeast transfer ribonucleic acid with sodium borohydride, J. Mol. Biol. 26, 55–66, 1967; Tanzer, M.L., Collagen reduction by sodium borohydride: effects of reconstitution, maturation, and lathyrism, Biochem. Biophys. Res. Commun. 32, 885–892, 1968; Phillips, T.M., Kosicki, G.W., and Schmidt, D.E., Jr., Sodium borohydride reduction of pyruvate by sodium borohydride catalyzed by pyruvate kinase, Biochim. Biophys. Acta 293, 125–133, 1973; Craig, A.S., Sodium borohydride as an aldehyde-blocking reagent for electron microscope histochemistry, Histochemistry 42, 141–144, 1974; Miles, E.W., Houck, D.R., and Floss, H.G., Stereochemistry of sodium borohydride reduction of tryptophan synthase of Escherichia coli and its amino acid Schiff’s bases, J. Biol. Chem. 257, 14203–14210, 1982; Kumar, A., Rao, P., and Pattabiraman, T.N., A colorimetric method for the estimation of serum glycated proteins based on differential reduction of free and bound glucose by sodium borohydride, Biochem. Med. Metab. Biol. 39, 296–304, 1988; Lenz, A.G., Costabel, U., Shaltiel, S., and Levine, R.L., Determination of carbonyl groups in oxidatively modified proteins by reduction with tritiated sodium borohydride, Anal. Biochem. 177, 419–425, 1989; Yan, L.J. and Sohal, R.S., Gel electrophoresis quantiation of protein carbonyls derivatized with tritiated sodium borohydride, Anal. Biochem. 265, 176–182, 1998; Azzam, T., Eliyahu, H., Shapira, L. et al., Polysaccharideoligoamine-based conjugates for gene delivery, J. Med. Chem. 45, 1817–1824, 2002; Purich, D.L., Use of sodium borohydride to detect acyl-phosphate linkages in enzyme reactions, Methods Enzymol. 354, 168–177, 2002; Bald, E., Chwatko, S., Glowacki, R., and Kusmierek, K., Analysis of plasma thiols by high-performance liquid chromatography with ultraviolet detection, J. Chromatog. A 1032, 109–115, 2004; Eike, J.H. and Palmer, A.F., Effect of NABH4 concentration and reaction time on physical properties of glutaraldehyde-polymerized hemoglobin, Biotechnol. Prog. 20, 946–952, 2004; Zhang, Z., Edwards, P.J., Roeske, R.W., and Guo, L., Synthesis and self-alkylation of isotope-coded affinity tag reagents, Bioconjug. Chem. 16, 458–464, 2005; Studelski, D.R., Giljum, K., McDowell, L.M., and Zhang, L., Quantitation of glycosaminoglycans by reversed-phase HPLC separation of fluorescent isoindole derivatives, Glycobiology 16, 65–72, 2006; Floor, E., Maples, A.M., Rankin, C.A. et al., A one-carbon modification of protein lysine associated with elevated oxidative stress in human substantia nigra, J. Neurochem. 97, 504–514, 2006; Kusmierek, K., Glowacki, R., and Bald, E., Analysis of urine for cysteine, cysteinylglycine, and homocysteine by high-performance liquid chromatography, Anal. BioAnal. Chem. 385, 855–860, 2006.

Sodium Chloride

Salt; NaCl


Ionic strength; physiological saline.

Sodium Cholate



Lindstrom, J., Anholt, R., Einarson, B. et al., Purification of acetylcholine receptors, reconstitution into lipid vesicles, and study of agonist-induced channel regulation, J. Biol. Chem. 255, 8340–8350, 1980; Gullick, W.J., Tzartos, S., and Lindstrom, J., Monoclonal antibodies as probes of acetylcholine receptor structure. 1. Peptide mapping, Biochemistry 20, 2173–2180, 1981; Henselman, R.A. and Cusanovich, M.A., The characterization of sodium cholate solubilized rhodopsin, Biochemistry 13, 5199–5203, 1974; Ninomiya, R., Masuoka, K., and Moroi, Y., Micelle formation of sodium chenodeoxycholate and solublization into the micelles: comparison with other unconjugated bile salts, Biochim. Biophys. Acta 1634, 116–125, 2003; Simoes, S.I., Marques, C.M., Cruz, M.E. et al., The effect of cholate on solubilization and permeability of simple and protein-loaded phosphatidylcholine/sodium cholate-mixed aggregates designed to mediate transdermal delivery of macromolecules, Eur. J. Pharm. Biopharm. 58, 509–519, 2004; Reis, S., Moutinho, C.G., Matos, C. et al., Noninvasive methods to determine the critical micelle concentration of some bile acid salts, Anal. Biochem. 334, 117–126, 2004; Nohara, D., Kajiura, T., and Takeda, K., Determination of micelle mass by electrospray ionization mass spectrometry, J. Mass Spectrom. 40, 489–493, 2005; Guo, J., Wu., T., Ping, Q. et al., Solublization and pharmacokinetic behaviors of sodium cholate/lecithin-mixed micelles containing cyclosporine A, Drug Deliv. 12, 35–39, 2005; Bottari, E., Buonfigli, A., and Festa, M.R., Composition of sodium cholate micellar solutions, Ann. Chim. 95, 479–490, 2005; Schweitzer, B., Felippe, A.C., Dal Bo, A. et al., Sodium dodecyl sulfate promoting a cooperative association process of sodium cholate with bovine serum albumin, J. Colloid Interface Sci. 298, 457–466, 2006; Burton, M.I., Herman, M.D., Alcain, F.J., and Villalba, J.M., Stimulation of polyprenyl 4-hydroxybenzoate transferase activity by sodium cholate and 3- [(cholamidopropyl)dimethylammonio]-1-propanesulfonate, Anal. Biochem. 353, 15–21, 2006; Ishibashi, A. and Nakashima, N., Individual dissolution of single-walled carbon nanotubes in aqueous solutions of steroid of sugar compounds and their Raman and near-IR spectral properties, Chemistry, 12, 7595–7602, 2006.

Sodium Cyanoborohydride

NaBH3 (CN)


Reducing agent; considered more selective than NaBH4.

Rosen, G.M., Use of sodium cyanoborohydride in the preparation of biologically active nitroxides, J. Med. Chem. 17, 358–360, 1974; Chauffe, L. and Friedman, M., Factors affecting cyanoborohydride reduction of aromatic Schiff’s bases in proteins, Adv. Exp. Med. Biol. 86A, 415–424, 1977; Baues, R.J. and Gray, G.R., Lectin purification on affinity columns containing reductively aminated disaccharides, J. Biol. Chem. 252, 57–60, 1977; Jentoft, N. and Dearborn, D.G., Labeling of proteins by reductive methylation using sodium cyanoborohydride, J. Biol. Chem. 254, 4359–4365, 1979; Jentoft, N., and Dearborn, D.G., Protein labeling by reductive methylation with sodium cyanoborohydride: effect of cyanide and metal ions on the reaction, Anal. Biochem. 106, 186–190, 1980; Bunn, H.F. and Higgins, P.T., Reaction of monosaccharides with proteins: possible evolutionary significance, Science 213, 222–224, 1981; Geoghegan, K.F., Cabacungan, J.C., Dixon, H.B., and Feeney, R.E., Alternative reducing agents for reductive methylation of amino groups in proteins, Int. J. Pept. Protein Res. 17, 345–352, 1981; Habeeb, A.F., Comparative studies on radiolabeling of lysozyme by iodination and reductive methylation, J. Immunol. Methods 65, 27–39, 1983; Prakash, C. and Vijay, I.K., A new fluorescent tag for labeling of saccharides, Anal. Biochem. 128, 41–46, 1983; Acharya, A.S. and Sussman, L.G., The reversibility of the ketoamine linkages of aldoses with proteins, J. Biol. Chem. 259, 4372–4378, 1984; Climent, I., Tsai, L., and Levine, R.L., Derivatization of gamma-glutamyl semialdehyde residues in oxidized proteins by fluorescamine, Anal. Biochem. 182, 226–232, 1989; Hartmann, C. and Klinman, J.P., Reductive trapping of substrate to methylamine oxidase from Arthrobacter P1, FEBS Lett. 261, 441–444, 1990; Meunier, F. and Wilkinson, K.J., Nonperturbing fluorescent labeling of polysaccharides, Biomacromolecules 3, 858–864, 2002; Webb, M.E., Stephens, E., Smith, A.G., and Abell, C., Rapid screening by MALDITOF mass spectrometry to probe binding specificity at enzyme active sites, Chem. Commun. 19, 2416–2417, 2003; Sando, S., Matsui, K., Niinomi, Y. et al., Facile preparation of DNA-tagged carbohydrates, Bioorg. Med. Chem. Lett. 13, 2633–2636, 2003; Peelen, D. and Smith, L.M., Immobilization of anine-modified oligonucleotides on aldehyde-terminated alkanethiol monolayers on gold, Langmuir 21, 266–271, 2005; Mirzaei, H. and Regnier, F., Enrichment of carbonylated peptides using Girard P reagent and strong cation exchange chromatography, Anal. Chem. 78, 770–778, 2006.

Sodium Deoxycholate

Desoxycholic Acid, Sodium Salt


Detergent; potential therapeutic use with adipose tissue.

Bril, C., van der Horst, D.J., Poort, S.R., and Thomas, J.B., Fractionation of spinach chloroplasts with sodium deoxycholate, Biochim. Biophys. Acta 172, 345–348, 1969; Smart, J.E. and Bonner, J., Selective dissociation of histones from chromatin by sodium deoxycholate, J. Mol. Biol. 58, 651–659, 1971; Part, M., Tarone, G., and Comoglio, P.M., Antigenic and immunogenic properties of membrane proteins solubilized by sodium desoxycholate, papain digestion, or high ionic strength, Immunochemistry 12, 9–17, 1975; Johansson, K.E. and Wbolewski, H., Crossed immunoelectrophoresis, in the presence of tween 20 or sodium deoxycholate, or purified membrane proteins from Acholeplasma laidlawii, J. Bacteriol. 136, 324–330, 1978; Lehnert, T. and Berlet, H.H., Selective inactivation of lactate dehydrogenase of rat tissues by sodium deoxycholate, Biochem. J. 177, 813–818, 1979; Suzuki, N., Kawashima, S., Deguchi, K., and Ueta, N., Low-density lipoproteins form human ascites plasma. Characterization and degradation by sodium deoxycholate, J. Biochem. 87, 1253–1256, 1980; Robern, H., The application of sodium deoxycholate and Sephacryl S-200 for the delipidation and separation of high-density lipoprotein, Experientia 38, 437–439, 1982; Nedivi, E. and Schramm, M., The beta-adrenergic receptor survives solubilization in deoxycholate while forming a stable association with the agonist, J. Biol. Chem. 259, 5803–5808, 1984; McKernan, R.M., Castro, S., Poat, J.A., and Wong, E.H., Solubilization of the N-methyl-D-aspartate receptor channel complex from rat and porcine brain, J. Neurochem. 52, 777–785, 1989; Carter, H.R. Wallace, M.A., and Fain, J.N., Activation of phospholipase C in rabbit brain membranes by carbachol in the presence of GTP gamma S: effects of biological detergents, Biochim. Biophys. Acta 1054, 129–134, 1990; Shivanna, B.D. and Rowe, E.S., Preservation of the native structure and function of Ca2+-ATPase from sarcoplasmic reticulum: solubilization and reconstitution by new short-chain phospholipid detergent 1,2-diheptanoyl-sn-phosphatidylcholine, Biochem. J. 325, 533–542, 1997; Arnold, U. and Ulbrich-Hofmann, R., Quantitative protein precipitation from guandine hydrochloride-containing solutions by sodium deoxycholate/trichloroacetic acid, Anal. Biochem. 271, 197–199, 1999; Haque, M.E., Das, A.R., and Moulik, S.P., Mixed micelles for sodium deoxycholate and polyoxyethylene sobitan monooleate (Tween 80), J. Colloid Interface Sci. 217, 1–7, 1999; Srivastava, O.P. and Srivastava, K., Characterization of a sodium deoxycholate-activable proteinase activity associated with betaA3/A1-crystallin of human lenses, Biochim. Biophys. Acta 1434, 331–346, 1999; Rotunda, A.M., Suzuki, H., Moy, R.L., and Kolodney, M.S., Detergent effects of sodium deoxycholate are a major feature of an injectable phosphatidylcholine formulation used for localized fat dissolution, Dermatol. Surg. 30, 1001–1008, 2004; Asmann, Y.W., Dong, M., and Miller, L.J., Functional characterization and purification of the secretin receptor expressed in baculovirus-infected insect cells, Regul. Pept. 123, 217–223, 2004; Ranganathan, R., Tcacenco, C.M., Rosseto, R., and Hajdu, J., Characterization of the kinetics of phospholipase C activity toward mixed micelles of sodium deoxycholate and dimyristoylphophatidylcholine, Biophys. Chem. 122, 79–89, 2006.


Shapiro, A.L., Vinuela, E., and Maizel, J.V., Jr., Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels, Biochem. Biophys. Res. Commun. 28, 815–820, 1967; Shapiro, A.L., and Maizel, J.V., Jr., Molecular weight estimation of polypeptides by SDS-polyacrylamide gel electrophoresis: further data concerning resolving power and general considerations, Anal. Biochem. 29, 505–514, 1969; Weber, K. and Osborn, M., The reliability of molecular weight determinations of dodecyl sulfate-polyacryalmide gel electrophoresis, J. Biol. Chem. 244, 4406–4412, 1969; Weber, K. and Kuter, D.J., Reversible denaturation of enzymes by sodium dodecyl sulfate, J. Biol. Chem. 246, 4504–4509, 1971; de Haen, C., Molecular weight standards for calibration of gel filtration and sodium dodecyl sulfatepolyacrylamide gel electrophoresis: ferritin and apoferritin, Anal. Biochem. 166, 235–245, 1987; Smith, B.J., SDS polyacrylamide gel electrophoresis of proteins, Methods Mol. Biol. 32, 23–34, 1994; Guttman, A., Capillary sodium dodecyl sulfate-gel electrophoresis of proteins, Electrophoresis 17, 1333–1341, 1996; Bischoff, K.M., Shi, L., and Kennelly, P.J., The detection of enzyme activity following sodium dodecyl sulfate-polyacryalamide gel electrophoresis, Anal. Biochem. 260, 1–17, 1998; Maizel, J.V., SDS polyacrylamide gel electrophoresis, Trends Biochem. Sci. 35, 590–592, 2000; Robinson, J.M. and Vandre, D.D, Antigen retrieval in cells and tissues: enhancement with sodium dodecyl sulfate, Histochem. Cell Biol. 116, 119–130, 2001; Todorov, P.D., Kralchevsky, P.A., Denkov, N.D. et al., Kinetics of solublization of n-decane and benzene by micellar solutions of sodium dodecyl sulfate, J. Colloid Interface Sci. 245, 371–382, 2002; Zhdanov, S.A., Starov, V.M., Sobolev, V.D., and Velarde, M.G., Spreading of aqueous SDS solutions over nitrocellulose membranes, J. Colloid Interface Sci. 264, 481–489, 2003; Santos, S.F., Zanette, D., Fischer, H., and Itri, R., A systematic study of bovine serum albumin (BSA) and sodium dodecyl sulfate (SDS) interactions by surface tension and small angle X-ray scattering, J. Colloid Interface Sci. 262, 400–408, 2003; Biswas, A. and Das, K.P., SDS-induced structural changes in alpha-crystallin and its effect on refolding, Protein J. 23, 529–538, 2004; Jing, P., Kaneta, T., and Imasaka, T., On-line concentration of a protein using denaturation by sodium dodecyl sulfate, Anal. Sci. 21, 37–42, 2005; Choi, N.S., Hahm, J.H., Maeng, P.J., and Kim, S.H., Comparative study of enzyme activity and stability of bovine and human plasmins in electrophoretic reagents, β-mercaptoethanol, DTT, SDS, Triton X-100, and urea, J. Biochem. Mol. Biol. 38, 177–181, 2005; Miles, A.P. and Saul, A., Quantifying recombinant proteins and their degradation products using SDS-PAGE and scanning laser densitometry, Methods Mol. Biol. 308, 349–356, 2005; Thongngam, M. and McClements, D.J., Influence of pH, ionic strength, and temperature on self-association and interactions of sodium dodecyl sulfate in the absence and presence of chitosan, Langmuir 21, 79–86, 2005; Romani, A.P., Gehlen, M.H., and Itri, R., Surfactant-polymer aggregates formed by sodium dodecyl sulfate, poly(N-vinyl-2-pyrrolidone), and poly(ethylene glycol), Langmuir 21, 1271–1233, 2005; Gudiksen, K.L., Gitlin, I., and Whitesides, G.M., Differentiation of proteins based on characteristic patterns of association and denaturation in solutions of SDS, Proc. Natl. Acad. Sci. USA 103, 7968–7972, 2006; Freitas, A.A., Paulo, L., Macanita, A.L, and Quina, F.H., Acid-base equilibria and dynamics in sodium dodecyl sulfate micelles: geminate recombination and effect of charge stabilization, Langmuir 22, 7986–7893, 2006.

Sodium Metabisulfite

Sodium Bisulfite


Mild reducing agent; converts unmethylated cytosine residues to uracil residues (DNA methylation).

Miller, R.F., Small, G., and Norris, L.C., Studies on the effect of sodium bisulfite on the stability of vitamin E, J. Nutr. 55, 81–95, 1955; Hayatsu, H., Wataya, Y., Kai, K., and Iida, S., Reaction of sodium bisulfite with uracil, cytosine, and their derivatives, Biochemistry 9, 2858–2865, 1970; Seno, T., Conversion of Escherichia coli tRNATrp to glutamine-accepting tRNA by chemical modification with sodium bisulfite, FEBS Lett. 51, 325–329, 1975; Tasheva, B. and Dessev, G., Artifacts in sodium dodecyl sulfate-polyacrylamide gel electrophoresis due to 2-mercaptoethanol, Anal. Biochem. 129, 98–102, 1983; Draper, D.E., Attachment of reporter groups to specific, selected cytidine residues in RNA using a bisulfite-catalyzed transamination reaction, Nucleic Acids Res. 12, 989–1002, 1984; Oakeley, E.J., DNA methylation analysis: a review of current methodologies, Pharmacol. Ther. 84, 389–400, 1999; Geisler, J.P., Manahan, K.J., and Geisler, H.E., Evaluation of DNA methylation in the human genome: why examine it and what method to use, Eur. J. Gynaecol. Oncol. 25, 19–24, 2004; Thomassin, H., Kress, C., and Grange, T., MethylQuant: a sensitive method for quantifying methylation of specific cytosines within the genome, Nucleic Acids Res. 32, e168, 2004; Derks, S., Lentjes, M.H., Mellebrekers, D.M. et al., Methylation-specific PCR unraveled, Cell. Oncol. 26, 291–299, 2004; Galm, O. and Herman, J.G., Methylation-specific polymerase chain reaction, Methods Mol. Biol. 113, 279–291, 2005; Ogino, S., Kawasaki, T., Brahmandam, M. et al., Precision and performance characteristics of bisulfite conversion and real-time PCR (MethylLight) for quantitative DNA methylation analysis, J. Mol. Diagn. 8, 209–217, 2006; Yang, I., Park, I.Y., Jang, S.M. et al., Rapid quantitation of DNA methylation through dNMP analysis following bisulfite PCR, Nucleic Acids Res. 34, e61, 2006; Wischnewski, F., Pantel, K., and Schwazenbach, H., Promoter demethylation and histone acetylation mediate gene expression of MAGE-A1, -A2, -A3, and -A12 in human cancer cells, Mol. Cancer Res. 4, 339–349, 2006; Zhou, Y., Lum, J.M., Yeo, G.H. et al., Simplified molecular diagnosis of fragile X syndrome by fluorescent methylation-specific PCR and GeneScan analysis, Clin. Chem. 52, 1492–1500, 2006.


Habeeb, A.F., Cassidy, H.G., and Singer, S.J., Molecular structural effects produced in proteins by reaction with succinic anhydride, Biochim. Biophys. Acta 29, 587–593, 1958; Hass, L.F., Aldolase dissociation into subunits by reaction with succinic anhydride, Biochemistry 3, 535–541, 1964; Scanu, A., Pollard, H., and Reader, W., Properties of human serum low-density lipoproteins after modification by succinic anhydride, J. Lipid Res. 9, 342–349, 1968; Vasilets, I.M., Moshkov, K.A., and Kushner, V.P., Dissociation of human ceruloplasmin into subunits under the action of alkali and succinic anhydride, Mol. Biol. 6, 193–199, 1972; Tedeschi, H., Kinnally, K.W., and Mannella, C.A., Properties of channels in mitochondrial outer membrane, J. Bioenerg. Biomembr. 21, 451–459, 1989; Palacian, E., Gonzalez, P.J., Pineiro, M., and Hernandez, F., Dicarboxylic acid anhydrides as dissociating agents of protein-containing structures, Mol. Cell. Biochem. 97, 101–111, 1990; Pavliakova, D., Chu, C., Bystricky, S. et al., Treatment with succinic anhydride improves the immunogenicity of Shigella flexneri type 2a O-specific polysaccharide-protein conjugates in mice, Infect. Immun. 67, 5526–5529, 1999; Ferretti, V., Gilli, P., and Gavezzotti, A., X-ray diffraction and molecular simulation study of the crystalline and liquid states of succinic anhydride, Chemistry 8, 1710–1718, 2002.



Osmolyte; density gradient centrifugation.

Cann, J.R., Coombs, R.O., Howlett, G.J. et al., Effects of molecular crowding on protein self-association: a potential source of error in sedimentation coefficients obtained by zonal ultracentrifugation in a sucrose gradient, Biochemistry 33, 10185–10190, 1994; Camacho-Vanegas, O., Lorein, F., and Amaldi, F., Flat absorbance background for sucrose gradients, Anal. Biochem. 228, 172–173, 1995; Ben-Zeev, O. and Doolittle, M.H., Determining lipase subunit structure by sucrose gradient centrifugation, Methods Mol. Biol. 109, 257–266, 1999; Lustig, A., Engel, A., Tsiotis, G. et al., Molecular weight determination of membrane proteins by sedimentation equilibrium at the sucrose of nycodenz-adjusted density of the hydrated detergent micelle, Biochim. Biophys. Acta 1464, 199–206, 2000; Kim, Y.S., Jones, L.A., Dong, A. et al., Effects of sucrose on conformational equilibria and fluctuations within the native-state ensemble of proteins, Protein Sci. 12, 1252–1261, 2003; Srinivas, K.A., Chandresekar, G., Srivastava, R., and Puvanakrishna, R., A novel protocol for the subcellular fractionation of C3A hepatoma cells using sucrose-density gradient centrifugation, J. Biochem. Biophys. Methods 60, 23–27, 2004; Richter, W., Determining the subunit structure of phosphodiesterase using gel filtration and sucrose-density gradient centrifugation, Methods Mol. Biol. 307, 167–180, 2005; Cioni, P., Bramanti, E., and Strambini, G.B., Effects of sucrose on the internal dynamics of azurin, Biophys. J. 88, 4213–4222, 2005; Desplats, P., Folco, E. and Salerno, G.L., Sucrose may play an additional role to that of an osmolyte in Synechocystis sp. PCC 6803 salt-shocked cells, Plant Physiol. Biochem. 43, 133–138, 2005; Chen, L., Ferreira, J.A., Costa, S.M. et al., Compaction of ribosomal protein S6 by sucrose occurs only under native conditions, Biochemistry 21, 2189–2199, 2006.


Good, N.E., Winget, G.D., Winter, W. et al., Hydrogen ion buffers for biological research, Biochemistry 5, 467–477, 1966; Itagaki, A. and Kimura, G., TES and HEPES buffers in mammalian cell cultures and viral studies: problem of carbon dioxide requirement, Exp. Cell Res. 83, 351–361, 1974; Bridges, S. and Ward, B., Effect of hydrogen ion buffers on photosynthetic oxygen evolution in the blue-green alga, Agmenellum quadruplicatum, Microbios 15, 49–56, 1976; Bailyes, E.M., Luzio, J.P., and Newby, A.C., The use of a zwitterionic detergent in the solubilization and purification of the intrinsic membrane protein 5′-nucleotidase, Biochem. Soc. Trans. 9, 140–141, 1981; Poole, C.A., Reilly, H.C., and Flint, M.H., The adverse effects of HEPES, TES, and BES zwitterionic buffers on the ultrastructure of cultured chick embryo epiphyseal chondrocytes, In Vitro 18, 755–765, 1982; Nakon, R. and Krishnamoorthy, C.R., Free-metal ion depletion by “Good’s” buffers, Science 221, 749–750, 1983; del Castillo, J., Escalona de Motta, G., Eterovic, V.A., and Ferchmin, P.A., Succinyl derivatives of N-Tris (hydroxylmethyl) methyl-2-aminoethane sulphonic acid: their effects on the frog neuromuscular junction, Br. J. Pharmacol. 84, 275–288, 1985; Kaushal, V. and Varnes, L.D., Effect of zwitterionic buffers on measurement of small masses of protein with bicinchoninic acid, Anal. Biochem. 157, 291–294, 1986; Bhattacharyya, A. and Yanagimachi, R., Synthetic organic pH buffers can support fertilization of guinea pig eggs, but not as efficiently as bicarbonate buffer, Gamete Res. 19, 123–129, 1988; Veeck, L.L., TES and Tris (TEST)-yolk buffer systems, sperm function testing, and in vitro fertilization, Fertil. Steril. 58, 484–486, 1992; Kragh-Hansen, U. and Vorum, H., Quantitative analyses of the interaction between calcium ions and human serum albumin, Clin. Chem. 39, 202–208, 1993; Jacobs, B.R., Caulfield, J., and Boldt, J., Analysis of TEST (TES and Tris) yolk buffer effects of human sperm, Fertil. Steril. 63, 1064–1070, 1995; Stellwagne, N.C., Bossi, A., Gelfi, C., and Righetti, P.G., DNA and buffers: are there any noninteracting, neutral pH buffers? Anal. Biochem. 287, 167–175, 2000; Taylor, J., Hamilton, K.L., and Butt, A.G., HCO3 - potentiates the cAMP-dependent secretory response of the human distal colon through a DIDS-sensitive pathway, Pflügers Arch. 442, 256–262, 2001; Taha, M., Buffers for the physiological pH range: acidic dissociation constants of zwitterionic compounds in various hydroorganic media, Ann. Chim. 95, 105–109, 2005.


Walseth, T.F., Graff, G., Moos, M.C., Jr., and Goldberg, N.D., Separation of 5′-ribonucleoside monophosphates by ion-pair reverse-phase high-performance liquid chromatography, Anal. Biochem. 107, 240–245, 1980; Ozkul, A. and Oztunc, A., Determination of naprotiline hydrochloride in tables by ion-pair extraction using bromthymol blue, Pharmzie 55, 321–322, 2000; Cecchi, T., Extended thermodynamic approach to ion interaction chromatography. Influence of the chain length of the solute ion; a chromatographic method for the determination of ion-pairing constants, J. Sep. Sci 28, 549–554, 2005; Pistos, C., Tsantili-Kakoulidou, A., and Koupparis, M., Investigation of the retention/pH profile of zwitterionic fluoroquinolones in reversed-phase and ion-interaction high-performance liquid chromatography, J. Pharm. Biomed. Anal. 39, 438–443, 2005; Choi, M.M., Douglas, A.D., and Murray, R.W., Ion-pair chromatographic separation of water-soluble gold monolayer-protected clusters, Anal. Chem. 78, 2779–2785, 2006; Saradhi, U.V., Prarbhakar, S., Reddy, T.J., and Vairamani, M., Ion-pair solid-phase extraction and gas chromatography mass spectrometric determination of acidic hydrolysis products of chemical warfare agents from aqueous samples, J. Chromatog. A, 1129, 9–13, 2006.


Leuty, S.J., Rapid dehydration of plant tissues for paraffin embedding; tetrahydrofuran vs. t-butanol, Stain Technol. 44, 103–104, 1969; Tandler, C.J. and Fiszer de Plazas, S., The use of tetrahydrofuran for delipidation and water solubilization of brain proteolipid proteins, Life Sci. 17, 1407–1410, 1975; Dressman, J.B., Himmelstein, K.J., and Higuchi, T., Diffusion of phenol in the presence of a complexing agent, tetrahydrofuran, J. Pharm. Sci. 72, 12–17, 1983; Diaz, R.S., Regueiro, P., Monreal, J., and Tandler, C.J., Selective extraction, solubilization, and reversed-phase high-performance liquid chromatography separation of the main proteins from myelin using tetrahydrofuran/water mixtures, J. Neurosci. Res. 29, 114–120, 1991; Santa, T., Koga, D., and Imai, K., Reversed-phase high-performance liquid chromatography of fullerenes with tetrahydrofuran-water as a mobile phase and sensitive ultraviolet or electrochemical detection, Biomed. Chromatogr. 9, 110–111, 1995; Lee, J., Kang, J.H., Lee, S.Y. et al., Protein kinase C ligands based on tetrahydrofuran templates containing a new set of phorbol ester pharmacophores, J. Med. Chem. 42, 4129–4139, 1999; Edwards, A.A., Ichihara, O., Murfin, S. et al., Tetrahydrofuran-based amino acids as library scaffolds, J. Comb. Chem. 6, 230–238, 2004; Baron, C.P., Refsgaard, H.H., Skibsted, L.H., and Andersen, M.L., Oxidation of bovine serum albumin initiated by the Fenton reaction — effect of EDTA, tert-butylhydroperoxide, and tetrahydrofuran, Free Radic. Res. 40, 409–417, 2006; Bowron, D.T., Finney, J.L., and Soper, A.K., The structure of liquid tetrahydrofuran, J. Am. Chem. Soc. 128, 5119–5126, 2006; Hermida, S.A., Possari, E.P., Souza, D.B. et al., 2′-deoxyguanosine, 2′-deoxycytidine, and 2′-deoxyadenosine adducts resulting from the reaction of tetrahydrofuran with DNA bases, Chem. Res. Toxicol. 19, 927–936, 2006; Li, A.C., Li, Y., Guirguis, M.S., Advantages of using tetrahydrofuran-water as mobile phases in the quantitation of cyclosporine A in monkey and rat plasma by liquid chromatography-tandem mass spectrometry, J. Pharm. Biomed. Anal. 43, 277–284, 2007.


Boxman, A.W., Barts, P.W., and Borst-Pauwels, G.W., Some characteristics of tetraphenylphosphonium uptake into Saccharomyces cerevisiae, Biochim. Biophys. Acta 686, 13–18, 1982; Flewelling, R.F. and Hubbell, W.L., Hydrophobic ion interactions with membranes. Thermodynamic analysis of tetraphenylphosphonium binding to vesicles, Biophys. J. 49, 531–540, 1986; Prasad, R. and Hofer, M., Tetraphenylphosphonium is an indicator of negative membrane potential in Candida albicans, Biochim. Biophys. Acta 861, 377–380, 1986; Aiuchi, T., Matsunada, M., Nakaya, K., and Nakamura, Y., Calculation of membrane potential in synaptosomes with use of a lipophilic cation (tetraphenylphosphonium), Chem. Pharm. Bull. 37, 3333–3337, 1989; Nhujak T. and Goodall, D.M., Comparison of binding of tetraphenylborate and tetraphenylphosphonium ion to cyclodextrins studied by capillary electrophoresis, Electrophoresis 22, 117–122, 2001; Yasuda, K., Ohmizo, C., and Katsu, T., Potassium and tetraphenylphosphonium ion-selective electrodes for monitoring changes in the permeability of bacterial outer and cytoplasmic membranes, J. Microbiol. Methods 54, 111–115, 2003; Min, J.J., Biswal, S., Deroose, C., and Gambhir, S.S., Tetraphenylphosphonium as a novel molecular probe for imaging tumors, J. Nucl. Med. 45, 636–643, 2004.


Rogers, D.R., Screening for amyloid with the thioflavin T fluorescent method, Am. J. Clin. Pathol. 44, 59–61, 1965; Saeed, S.M. and Fine, G., Thioflavin T for amyloid, Am. J. Clin. Pathol. 47, 588–593, 1967; Levine, H., III, Stopped-flow kinetics reveal multiple phase of thioflavin T binding to Alzheimer beta (1–40) amyloid fibrils, Arch. Biochem. Biophys. 342, 306–316, 1997; De Ferrari, G.V., Mallender, W.D., Inestrosa, N.C., and Rosenberry, T.L., Thioflavin T is a fluorescent probe of the acetylcholinesterase peripheral site that reveals conformational interactions between the peripheral and acylation sites, J. Biol. Chem. 276, 23282–23287, 2001; Ban, T., Hamada, D., Hasegawa, K. et al., Direct observation of amyloid fibril growth monitored by thioflavin T fluorescence, J. Biol. Chem. 278, 16462–16465, 2003; Krebs, M.R., Bromley, E.H., and Donald, A.M., The binding of thioflavin T to amyloid fibrils: localization and implications, J. Struct. Biol. 149, 30–37, 2005; Khurana, R., Coleman, C., Ionescu-Zanetti, C. et al., Mechanisms of thioflavin T binding to amyloid fibrils, J. Struct. Biol. 151, 229–238, 2005; Darhal, N., Garnier-Suillerot, A., and Salerno, M., Mechanism of thioflavin T accumulation inside cells overexpressing P-glycoprotein or multidrug resistance-associated protein: role of lipophilicity and positive charge, Biochem. Biophys. Res. Commun. 343, 623–629, 2006; Eisert, R., Felau, L., and Brown, L.R., Methods for enhancing the accuracy and reproducibility of Congo red and thioflavin T assays, Anal. Biochem. 353, 144–146, 2006.

Thionyl Chloride

Sulfurous Oxychloride


Preparation of acyl chlorides.

Rodin, R.L. and Gershon, H., Photochemical alpha-chlorination of fatty acid chlorides by thionyl chloride, J. Org. Chem. 38, 3919–3921, 1973; DuVal, G., Swaisgood, H.E., and Horton, H.R, Preparation and characterization of thionyl chloride-activated succinamidopropyl-glass as a covalent immobilization matrix, J. Appl. Biochem. 6, 240–250, 1984; Molnar-Perl, I., Pinter-Szakacs, M., and Fabian-Vonsik, V., Esterification of amino acids with thionyl chloride acidified butanols for their gas chromatographic analysis, J. Chromatog. 390, 434–438, 1987; Stabel, T.J., Casele, E.S., Swaisgood, H.E., and Horton, H.R., Anti-IgG immobilized controlled pore glass. Thionyl chloride-activated succinamidopropyl-gas as a covalent immobization matrix, Appl. Biochem. Biotechnol. 36, 87–96, 1992; Chamoulaud, G. and Belanger, D., Chemical modification of the surface of a sulfonated membrane by formation of a sulfonamide bond, Langmuir 20, 4989–4895, 2004; Porjazoska, A.,Yilmaz, O.K., Baysal, K. et al., Synthesis and characterization of poly(ethylene glycol)-poly(D,L-lactide-co-glycolide) poly(ethylene glycol) tri-block co-polymers modified with collagen: a model surface suitable for cell interaction, J. Biomater. Sci. Polym. Ed. 17, 323–340, 2006; Gao, C., Jin, Z.Q., Kong, H. et al., Polyureafunctionalized multiwalled carbon nanotubes: synthesis, morphology, and Ramam spectroscopy, J. Phys. Chem. B 109, 11925–11932, 2005; Chen, G.X., Kim, H.S., Park, B.H., and Yoon, J.S., Controlled functionalization of multiwalled carbon nanotubes with various molecular-weight poly(L-lactic acid), J. Phys. Chem. B 109, 22237–22243, 2005.


Maloof, F. and Soodak, M., Cleavage of disulfide bonds in thyroid tissue by thiourea, J. Biol. Chem. 236, 1689–1692, 1961; Gerfast, J.A., Automated analysis for thiourea and its derivatives in biological fluids, Anal. Biochem. 15, 358–360, 1966; Lippe, C., Urea and thiourea permeabilities of phospholipid and cholesterol bilayer membranes, J. Mol. Biol. 39, 588–590, 1966; Carlsson, J., Kierstan, M.P., and Brocklehurst, K., Reactions of L-ergothioneine and some other aminothiones with 2,2′- and 4,4′-dipyridyl disulphides and of L-ergothioneine with iodoacetamide, 2-mercaptoimidazoles, and 4-thioypyridones, thiourea, and thioacetamide as highly reactive neutral sulphur nucleophiles, Biochem. J. 139, 221–235, 1974; Filipski, J., Kohn K.W., Prather, R., and Bonner, W.M., Thiourea reverses crosslinks and restores biological activity in DNA treated with dichlorodiaminoplatinum (II), Science 204, 181–183, 1979; Wasil, M., Halliwell, B., Grootveld, M. et al., The specificity of thiourea, dimethylthiourea, and dimethyl sulphoxide as scavengers of hydroxyl radicals. Their protection of alpha-1-antiproteinase against inactivation by hypochlorous acid, Biochem. J. 243, 867–870, 1987; Doona, C.J. and Stanbury, D.M., Equilibrium and redox kinetics of copper(II)–thiourea complexes, Inorg. Chem. 35, 3210–3216, 1996; Rabilloud, T., Use of thiourea to increase the solubility of membrane proteins in two-dimensional electrophoresis, Electrophoresis 19, 758–760, 1998; Musante, L., Candiano, G., and Ghiggeri, G.M., Resolution of fibronectin and other uncharacterized proteins by two-dimensional polyacrylamide electrophoresis with thiourea, J. Chromatog. B 705, 351–356, 1998; Nagy, E., Mihalik, R., Hrabak, A. et al., Apoptosis inhibitory effect of the isothiourea compound, tri-(2-thioureido-S-ethyl)-amine, Immunopharmacology 47, 25–33, 2000; Galvani, M., Rovatti, L., Hamdan, M. et al., Protein alkylation in the presence/absence of thiourea in proteome analysis: a matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry investigation, Electrophoresis 22, 2066–2074, 2001; Castellanos-Serra, L. and Paz-Lago, D., Inhibition of unwanted proteolysis during sample preparation: evaluation of its efficiency in challenge experiments, Electrophoresis 23, 1745–1753, 2002; Tyagarajan, K., Pretzer, E., and Wiktorowicz, J.E., Thiol-reactive dyes for fluorescence labeling of proteomic samples, Electrophoresis 24, 2348–2358, 2003; Fuerst, D.E., and Jacosen, E.N., Thiourea-catalyzed enantioselective cyanosilylation of ketones, J. Am. Chem. Soc. 127, 8964–8965, 2005; Gomez, D.E., Fabbrizzi, L., Licchelli, M., and Monzani, E., Urea vs. thiourea in anion recognition, Org. Biomol. Chem. 3, 1495–1500, 2005; George, M., Tan, G., John, V.T., and Weiss, R.G., Urea and thiourea derivatives as low molecular-mass organochelators, Chemistry 11, 3243–3254, 2005; Limbut, W., Kanatharana, P., Mattiasson, B. et al., A comparative study of capacitive immunosensors based on self-assembled monolayers formed from thiourea, thioctic acid, and 3-mercaptopropionic acid, Biosens. Bioelectron. 22, 233–240, 2006.


Habeeb, A.F., Determination of free amino groups in proteins by trinitrobenzenesulfonic acid, Anal. Biochem. 14, 328–336, 1966; Goldfarb, A.R., A kinetic study of the reactions of amino acids and peptides with trinitrobenzenesulfonic acid, Biochemistry 5, 2570–2574, 1966; Scheele, R.B. and Lauffer, M.A., Restricted reactivity of the epsilon-amino groups of tobacco mosaic virus protein toward trinitrobenzenesulfonic acid, Biochemistry 8, 3597–3603, 1969; Godin, D.V. and Ng, T.W., Trinitrobenzenesulfonic acid: a possible chemical probe to investigate lipid–protein interactions in biological membranes, Mol. Pharmacol. 8, 426–437, 1972; Bubnis, W.A. and Ofner, C.M., III, The determination of epsilon-amino groups in soluble and poorly soluble proteinaceous materials by a spectrophotometric method using trinitrobenzenesulfonic acid, Anal. Biochem. 207, 129–133, 1992; Cayot, P. and Tainturier, G., The quantification of protein amino groups by the trinitrobenzenesulfonic acid method: a reexamination, Anal. Biochem. 249, 184–200, 1997; Neurath, M., Fuss, I., and Strober, W., TNBS-colitis, Int. Rev. Immunol. 19, 51–62, 2000; Lindsay, J., Van Montfrans, C., Brennen, F. et al., IL-10 gene therapy prevents TNBS-induced colitis, Gene Ther. 9, 1715–1721, 2002; Whittle, B.J., Cavicchi, M., and Lamarque, D., Assessment of anticolitic drugs in the trinitrobenzenesulfonic acid (TNBS) rat model of inflammatory bowel disease, Methods Mol. Biol. 225, 209–222, 2003; Necefli, A., Tulumoglu, B., Giris, M. et al., The effects of melatonin on TNBS-induced colitis, Dig. Dis. Sci. 51, 1538–1545, 2006.


Sokolovsky, M., Riordan, J.F., and Vallee, B.L., Tetranitromethane. A reagent for the nitration of tyrosyl residues in proteins, Biochemistry 5, 3582–3589, 1966; Nishikimi, M. and Yagi, K., Reaction of reduced flavins with tetranitromethane, Biochem. Biophys. Res. Commun. 45, 1042–1048, 1971; Kunkel, G.R., Mehrabian, M., and Martinson, H.G., Contact-site crosslinking agents, Mol. Cell. Biochem. 34, 3–13, 1981; Rial, E. and Nicholls, D.G., Chemical modification of the brown-fatmitochondrial uncoupling protein with tetranitromethane and N-ethylmaleimide. A cysteine residue is implicated in the nucleotide regulation of anion permeability, Eur. J. Biochem. 161, 689–694, 1986; Prozorovski, V., Krook, M., Atrian, S. et al., Identification of reactive tyrosine residues in cysteine-reactive dehydrogenases. Differences between liver sorbitol, liver alcohol, and Drosophila alcohol dehydrogenase, FEBS Lett. 304, 46–50, 1992; Gadda, G., Banerjee, A., and Fitzpatrick, P.F., Identification of an essential tyrosine residue in nitroalkane oxidase by modification with tetranitromethane, Biochemistry 39, 1162–1168, 2000; Hodges, G.R. and Ingold, K.U., Superoxide, amine buffers, and tetranitro-methane: a novel free radical chain reaction, Free Radic. Res. 33, 547–550, 2000; Capeillere-Blandin, C., Gausson, V., DescampsLatscha, B., and Witko-Sarsat, V., Biochemical and spectrophotometric significance of advanced oxidation protein products, Biochim. Biophys. Acta 1689, 91–102, 2004; Lundblad, R.L., Chemical Reagents for Protein Modification, CRC Press, Boca Raton, FL, 2004; Negrerie, M., Martin, J.L., and Nghiem, H.O., Functionality of nitrated acetylcholine receptor: the two-step formation of nitrotyrosines reveals their differential role in effectors binding, FEBS Lett. 579, 2643–2647, 2005; Carven, G.J. and Stern, L.J., Probing the ligand-induced conformational change in HLA-DR1 by selective chemical modification and mass spectrometry mapping, Biochemistry 44, 13625–13637, 2005.


α-D-glucopyranoglucopyranosyl-1,1-α-Dglucopyranoside; Mycose


A nonreducing sugar that is found in a variety of organisms where it is thought to protect against stress such as dehydration; there is considerable interest in the use of trehalose as a stabilizer in biopharmaceutical proteins.

Elbein, A.D., The metabolism of alpha, alpha-trehalose, Adv. Carbohydr. Chem. Biochem. 30, 227–256, 1974; Wiemken, A., Trehalose in yeast, stress protectant rather than reserve carbohydrate, Antonie Van Leeuwenhoek, 58, 209–217, 1990; Newman, Y.M., Ring, S.G., and Colaco, C., The role of trehalose and other carbohydrates in biopreservation, Biotechnol. Genet. Eng. Rev. 11, 263–294, 1993; Panek, A.D., Trehalose metabolism — new horizons in technological applications, Braz. J. Med. Biol. Res. 28, 169–181, 1995; Schiraldi, C., Di Lernia, I., and De Rosa, M., Trehalose production: exploiting novel approaches, Trends Biotechnol. 20, 420–425, 2002; Elbein, A.D., Pan, Y.T., Pastuszak, I., and Carroll, D., New insights on trehalose: a multifunctional molecule, Glycobiology 13, 17R–27R, 2003; Gancedo, C. and Flores, C.L., The importance of a functional trehalose biosynthetic pathway for the life of yeasts and fungi, FEMS Yeast Res. 4, 351–359, 2004; Cordone, L., Cottone, G., Giuffrida, S. et al., Internal dynamics and protein-matrix coupling in trehalose-coated proteins, Biochim. Biophys. Acta 1749, 252–281, 2005.


Chang, Y.C., Efficient precipitation and accurate quantitation of detergent-solubilized membrane proteins, Anal. Biochem. 205, 22–26, 1992; Sivaraman, T., Kumar, T.K., Jayaraman, G., and Yu. C., The mechanism of 2,2,2-trichloroacetic acid-induced protein precipitation, J. Protein Chem. 16, 291–297, 1997; Arnold, U. and Ulbrich-Hoffman, R., Quantitative protein precipation form guandine hydrochloride-containing solutions by sodium deoxycholate/trichloroacetic acid, Anal. Biochem. 271, 197–199, 1999; Jacobs, D.I., van Rijssen, M.S., van der Heijden, R., and Verpoorte, R., Sequential solubilization of proteins precipitated with trichloroacetic acid in acetone from cultured Catharanthus roseus cells yields 52% more spots after two-dimensional electrophoresis, Proteomics 1, 1345–1350, 2001; Garcia-Rodriguez, S., Castilla, S.A., Machado, A., and Ayala, A., Comparison of methods for sample preparation of individual rat cerebrospinal fluid samples prior to two-dimensional polyacrylamide gel electrophoresis, Biotechnol. Lett. 25, 1899–1903, 2003; Chen, Y.Y., Lin, S.Y., Yeh, Y.Y. et al., A modified protein precipitation procedure for efficient removal of albumin from serum, Electrophoresis 26, 2117–2127, 2005; Zellner, M., Winkler, W., Hayden, H. et al., Quantitative validation of different protein precipitation methods in proteome analysis of blood platelets, Electrophoresis 26, 2481–2489, 2005; Carpentier, S.C., Witters, E., Laukens, K. et al., Preparation of protein extracts from recalcitrant plant tissues: an evaluation of different methods for two-dimensional gel electrophoresis analysis, Proteomics 5, 2497–2507, 2005; Manadas, B.J., Vougas, K., Fountoulakis, M., and Duarte, C.B., Sample sonication after trichloroacetic acid precipitation increases protein recovery from cultured hippocampal neurons, and improves resolution and reproducibility in two-dimensional gel electrophoresis, Electrophoresis 27, 1825–1831, 2006; Wang, A., Wu, C.J., and Chen, S.H., Gold nanoparticle-assisted protein enrichment and electroelution for biological samples containing low protein concentration — a prelude of gel electrophoresis, J. Proteome Res. 5, 1488–1492, 2006.


Fitzgerald, J.W., The Tris-catalyzed isomerization of potassium D-glucose 6-O-sulfate, Can. J. Biochem. 53, 906–910, 1975; Buhl, S.N., Jackson, K.Y., and Graffunder, B., Optimal reaction conditions for assaying human lactate dehydrogenase pyruvate-to-lactate at 25, 30, and 37 degrees C, Clin. Chem. 24, 261–266, 1978; Myohanen, T.A., Bouriotas, V., and Dean, P.D., Affinity chromatography of yeast alpha-glucosidase using ligand-mediated chromatography on immobilized phenylboronic acids, Biochem. J. 197, 683–688, 1981; Shinomiya, Y., Kato, N., Imazawa, M., and Miyamoto, K., Enzyme immunoassay of the myelin basic protein, J. Neurochem. 39, 1291–1296, 1982; Arita, M., Iwamori, M., Higuchi, T., and Nagai, Y., 1,1,3,3-tetramethylurea and triethanolaminme as a new useful matrix for fast atom bombardment mass spectrometry of gangliosides and neutral glycosphingolipids, J. Biochem. 93, 319–322, 1983; Cao, H. and Preiss, J., Evidence for essential arginine residues at the active site of maize branching enzymes, J. Protein Chem. 15, 291–304, 1996; Knaak, J.B., Leung, H.W., Stott, W.T. et al., Toxicology of mono-, di-, and triethanolamine, Rev. Environ. Contim. Toxicol. 149, 1–86, 1997; Liu, Q., Li, X., and Sommer, S.S., pK-matched running buffers for gel electrophoresis, Anal. Biochem. 270, 112–122, 1999; Sanger-van de Griend, C.E., Enantiomeric separation of glycyl dipeptides by capillary electrophoresis with cyclodextrins as chiral selectors, Electrophoresis 20, 3417–3424, 1999; Fang, L., Kobayashi, Y., Numajiri, S. et al., The enhancing effect of a triethanolamine-ethanol-isopropyl myristate mixed system on the skin permeation of acidic drugs, Biol. Pharm. Bull. 25, 1339–1344, 2002; Musial, W. and Kubis, A., Effect of some anionic polymers of pH of triethanolamine aqueous solutions, Polim. Med. 34, 21–29, 2004.


Brind, J.L., Kuo, S.W., Chervinsky, K., and Orentreich, N., A new reversed-phase, paired-ion thin-layer chromatographic method for steroid sulfate separations, Steroids 52, 561–570, 1988; Koves, E.M., Use of high-performance liquid chromatography-diode array detection in forensic toxicology, J. Chromatog. A 692, 103–119, 1995; Cole, S.R. and Dorsey, J.G., Cyclohexylamine additives for enhanced peptide separations in reversed-phase liquid chromatography, Biomed. Chromatog. 11, 167–171, 1997; Gilar, M. and Bouvier, E.S.P., Purification of crude DNA oligonucleotides by solid-phase extraction and reversed-phase high-performance liquid chromatography, J. Chromatog. A 890, 167–177, 2000; Loos, R. and Barcelo, D., Determination of haloacetic acids in aqueous environments by solid-phase extraction followed by ion-pair liquid chromatography-electrospray ionization mass spectrometric detection, J. Chromatog. A 938, 45–55, 2001; Gilar, M., Fountain, K.J., Budman, Y. et al., Ion-pair reversed-phase high-performance liquid chromatography analysis of oligonucleotides: retention prediction, J. Chromatog. A 958, 167–182, 2002; El-dawy, M.A., Mabrouk, M.M., and ElBarbary, F.A., Liquid chromatographic determination of fluoxetine, J. Pharm. Biomed. Anal. 30, 561–571, 2002; Yang, X., Zhang, X., Li, A. et al., Comprehensive two-dimensional separations based on capillary high-performance liquid chromatography and microchip electrophoresis, Electrophoresis 24, 1451–1457, 2003; Murphey, A.T., Brown-Augsburger, P., Yu, R.Z. et al., Development of an ion-pair reverse-phase liquid chromatographic/tandem mass spectrometry method for the determination of an 18-mer phosphorothioate oligonucleotide in mouse liver tissue, Eur. J. Mass Spectrom. 11, 209–215, 2005; Xie, G., Sueishi, Y., and Yamamoto, S., Analysis of the effects of protic, aprotic, and multi-component solvents on the fluorescence emission of naphthalene and its exciplex with triethylamine, J. Fluoresc. 15, 475–483, 2005.

Trifluoroacetic Acid


Ion-pair reagent; HLPC; peptide synthesis.

Rosbash, D.O. and Leavitt, D., Decalcification of bone with trifluoroacetic acid, Am. J. Clin. Pathol. 22, 914–915, 1952; Katz, J.J., Anhydrous trifluoroacetic acid as a solvent for proteins, Nature 174, 509, 1954; Uphaus, R.A., Grossweiner, L.I., Katz, J.J., and Kopple, K.D., Fluorescence of tryptophan derivatives in trifluoroacetic acid, Science 129, 641–643, 1959; Acharya, A.S., di Donato, A., Manjula, B.N. et al., Influence of trifluoroacetic acid on retention times of histidine-containing tryptic peptides in reverse phase HPLC, Int. J. Pept. Protein Res. 22, 78–82, 1983; Tsugita, A., Uchida, T., Mewes, H.W., and Ataka, T., A rapid vapor-phase acid (hydrochloric and trifluoroacetic acid) hydrolysis of peptide and protein, J. Biochem. 102, 1593–1597, 1987; Hulmes, J.D. and Pan, Y.C., Selective cleavage of polypeptides with trifluoroacetic acid: applications for microsequencing, Anal. Biochem. 197, 368–376, 1991; Eshragi, J. and Chowdhury, S.K., Factors affecting electrospray ionization of effluents containing trifluoroacetic acid for high-performance liquid chromatography/mass spectrometry, Anal. Chem. 65, 3528–3533, 1993; Apffel, A., Fischer, S., Goldberg, G. et al., Enhanced sensitivity for peptide mapping with electrospray liquid chromatography-mass spectrometry in the presence of signal suppression due to trifluoroacetic acid-containing mobiles phases, J. Chromatog. A 712, 177–190, 1995; Guy, C.A. and Fields, G.B., Trifluoroacetic acid cleavage and deprotection of resin-bound peptides following synthesis by Fmoc chemistry, Methods Enzymol. 289, 67–83, 1997; Morrison, I.M. and Stewart, D., Plant cell wall fragments released on solubilization in trifluoroacetic acid, Phytochemistry 49, 1555–1563, 1998; Yan, B., Nguyen, N., Liu, L. et al., Kinetic comparison of trifluoroacetic acid cleavage reactions of resin-bound carbamates, ureas, secondary amides, and sulfonamides from benzyl-, benzhyderyl-, and indole-based linkers, J. Comb. Chem. 2, 66–74, 2000; Ahmad, A., Madhusudanan, K.P., and Bhakuni, V., Trichloroacetic acid- and trifluoroacetic acid-induced unfolding of cytochrome C: stabilization of a nativelike fold intermediate(1), Biochim. Biophys. Acta 1480, 201–210, 2000; Chen, Y., Mehok, A.R., Mant, C.T. et al., Optimum concentration of trifluoroacetic acid for reversed-phase liquid chromatography of peptide revisited, J. Chromatog. A 1043, 9–18, 2004.


Bernhard, S.A., Ionization constants and heats of Tris(hydroxymethyl)aminomethane and phosphate buffers, J. Biol. Chem. 218, 961–969, 1956; Rapp, R.D. and Memminger, M.M., Tris(hydroxymethyl)aminomethane as an electrophoresis buffer, Am. J. Clin. Pathol. 31, 400–403, 1959; Rodkey, F.L., Tris(hydroxymethyl)aminomethane as a standard for Kjeldahl nitrogen analysis, Clin. Chem. 10, 606–610, 1964; Oliver, R.W. and Viswanatha, T., Reaction of Tris(hydroxymethyl)aminomethane with cinnamoyl imidazole and cinnamoyltrypsin, Biochim. Biophys. Acta 156, 422–425, 1968; Douzou, P., Enzymology at subzero temperatures, Mol. Cell. Biochem. 1, 15–27, 1973; Fitzgerald, J.W., The Tris-catalyzed isomerization of potassium D-glucose 6-O-sulfate, Can. J. Biochem. 53, 906–910, 1975; Visconti, M.A. and Castrucci, A.M., Tris buffer effects on melanophore-aggegrating responses, Comp. Biochem. Physiol. C 82, 501–503, 1985; Stambler, B.S., Grant, A.O., Broughton, A., and Strauss, H.C., Influences of buffers on dV/dtmax recovery kinetics with lidocaine in myocardium, Am. J. Physiol. 249, H663–H671, 1985; Nakano, M. and Tauchi, H., Difference in activation by Tris(hydroxymethyl)aminomethane of Ca,Mg-ATPase activity between young and old rat skeletal muscles, Mech. Aging Dev. 36, 287–294, 1986; Oliveira, L., Araujo-Viel, M.S., Juliano, L., and Prado, E.S., Substrate activation of porcine kallikrein N- α derivatives of arginine 4-nitroanilides, Biochemistry 26, 5032–5035, 1987; Ashworth, C.D. and Nelson, D.R., Antimicrobial potentiation of irrigation solutions containing Tris-[hydroxymethyl] aminomethane-EDTA, J. Am. Vet. Med. Assoc. 197, 1513–1514, 1990; Schacker, M., Foth, H., Schluter, J., and Kahl, R., Oxidation of Tris to one-carbon compounds in a radical-producing model system, in microsomes, in hepatocytes, and in rats, Free Radic. Res. Commun. 11, 339–347, 1991; Weber, R.E., Use of ionic and zwitterionic (Tris/BisTris and HEPES) buffers in studies on hemoglobin function, J. Appl. Physiol. 72, 1611–1615, 1992; Veeck, L.L., TES and Tris (TEST)-yolk buffer systems, sperm function testing, and in vitro fertilization, Fertil. Steril. 58, 484–486, 1992; Shiraishi, H., Kataoka, M., Morita, Y., and Umemoto, J., Interaction of hydroxyl radicals with Tris (hydroxymethyl)aminomethane and Good’s buffers containing hydroxymethyl or hydroxyethyl residues produce formaldehyde, Free Radic. Res. Commun. 19, 315–321, 1993; Vasseur, M., Frangne, R., and Alvarado, F., Buffer-dependent pH sensitivity of the fluorescent chloride-indicator dye SPQ, Am. J. Physiol. 264, C27–C31, 1993; Niedernhofer, L.J., Riley, M., Schnez-Boutand, N. et al., Temperature-dependent formation of a conjugate between Tris(hydroxymethyl) aminomethane buffer and the malondialdehyde-DNA adduct pyrimidopurinone, Chem. Res. Toxicol. 10, 556–561, 1997; Trivic, S., Leskovac, V., Zeremski, J. et al., Influence of Tris(hydroxymethyl)aminomethane on kinetic mechanism of yeast alcohol dehydrogenase, J. Enzyme Inhib. 13, 57–68, 1998; Afifi, N.N., Using difference spectrophotometry to study the influence of different ions and buffer systems on drug protein binding, Drug Dev. Ind. Pharm. 25, 735–743, 1999; AbouHaider, M.G. and Ivanov, I.G., Nonenzymatic RNA hydrolysis promoted by the combined catalytic activity of buffers and magnesium ions, Z. Naturforsch. 54, 542–548, 1999; Shihabi, Z.K., Stacking of discontinuous buffers in capillary zone electrophoresis, Electrophoresis 21, 2872–2878, 2000; Stellwagen, N.C, Bossi, A., Gelfi, C., and Righetti, P.G., DNA and buffers: are there any noninteracting, neutral pH buffers? Anal. Biochem. 287, 167–175, 2000; Burcham, P.C., Fontaine, F.R., Petersen, D.R., and Pyke, S.M., Reactivity of Tris(hydroxymethyl) aminomethane confounds immunodetection of acrolein-adducted proteins, Chem. Res. Toxicol. 16, 1196–1201, 2003; Koval, D., Kasicka, V., and Zuskova, I., Investigation of the effect of ionic strength of Tris-acetate background electrolyte on electrophoretic mobilities of mono-, di-, and trivalent organic anions by capillary electrophoresis, Electrophoresis 26, 3221–3231, 2005; Kinoshita, T., Yamaguchi, A., and Tada, T., Tris(hydroxymethyl)aminomethane-induced conformational change and crystal-packing contraction of porcine pancreatic elastase, Acta Crystallograph. Sect. F Struct. Biol. Cryst. Commun. 62, 623–626, 2006; Qi, Z., Li, X., Sun, D. et al., Effect of Tris on catalytic activity of MP-11, Bioelectrochemistry 68, 40–47, 2006.


Gray, W.R., Disulfide structures of highly bridged peptides: a new strategy for analysis, Protein Sci. 2, 1732–1748, 1993; Gray, W.R., Echistatin disulfide bridges: selective reduction and linkage assignment, Protein Sci. 2, 1749–1755, 1993; Han, J.C. and Han, G.Y., A procedure for quantitative determination of Tris(2-carboxyethyl)phosphine, an odorless reducing agent more stable and effective than dithiothreitol, Anal. Biochem. 220, 5–10, 1994; Wu, J., Gage, D.A., and Watson, J.T., A strategy to locate cysteine residues in proteins by specific chemical cleavage followed by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry, Anal. Biochem. 235, 161–174, 1996; Han, J., Yen. S., Han, G., and Han, F., Quantitation of hydrogen peroxide using Tris(2-carboxyethyl) phosphine, Anal. Biochem. 234, 107–109, 1996; Han, J., Clark, C., Han, G. et al., Preparation of 2-nitro-5-thiobenzoic acid using immobilized Tris(2-carboxyethyl) phosphine, Anal. Biochem. 268, 404–407, 1999; Anderson, M.T., Trudell, J.R., Voehringer, D.W. et al., An improved monobromobimane assay for glutathione utilizing Tris-(2-carboxyethyl)phosphine as the reductant, Anal. Biochem. 272, 107–109, 1999; Shafer, D.E., Inman, J.K. and Lees, A. Reaction of Tris(2-carboxyethyl)phosphine (TCEP) with maleimide and alpha-haloacyl groups: anomalous elution of TCEP by gel filtration, Anal. Biochem. 282, 161–164, 2000; Rhee, S.S. and Burke, D.H., Tris(2-carboxyethyl)phosphine stabilization of RNA: comparison with dithiothreitol for use with nucleic acid and thiophosphoryl chemistry, Anal. Biochem. 325, 137–143, 2004; Legros, C., Celerier, M.L., and Guette, C., An unusual cleavage reaction of a peptide observed during dithiothreitol and Tris(2-carboxyethyl)phosphine reduction: application to sequencing of HpTx2 spider toxin using nanospray tandem mass spectrometry, Rapid Commun. Mass Spectrom. 19, 1317–1323, 2004; Xu, G., Kiselar, J., He, Q., and Chance, M.R., Secondary reactions and strategies to improve quantitative protein footprinting, Anal. Chem. 77, 3029–3037, 2005; Valcu, C.M. and Schlink, K., Reduction of proteins during sample preparation and two-dimensional gel electrophoresis of woody plant samples, Proteomics 6, 1599–1605, 2006; Scales, C.W., Convertine, A.J., and McCormick, C.L., Fluorescent labeling of RAFT-generated poly(N-isopropylacrylamide) via a facile maleimide-thiol coupling reaction, Biomacromolecules 7, 1389–1392, 2006.




Chaotropic agent.

Edelhoch, H., The effect of urea analogues and metals on the rate of pepsin denaturation, Biochim. Biophys. Acta 22, 401–402, 1956; Steven, F.S. and Tristram, G.R., The denaturation of ovalbumin. Changes in optical rotation, extinction, and viscosity during serial denaturation in solution of urea, Biochem. J. 73, 86–90, 1959; Nelson, C.A. and Hummel, J.P., Reversible denaturation of pancreatic ribonuclease by urea, J. Biol. Chem. 237, 1567–1574, 1962; Herskovits, T.T., Nonaqueous solutions of DNA; denaturation by urea and its methyl derivatives, Biochemistry 2, 335–340, 1963; Subramanian, S., Sarma, T.S., Balasubramanian, D., and Ahluwalia, J.C., Effects of the urea–guanidinium class of protein denaturation on water structure: heats of solution and proton chemical shift studies, J. Phys. Chem. 75, 815–820, 1971; Strachan, A.F., Shephard, E.G., Bellstedt, D.U. et al., Human serum amyloid A protein. Behavior in aqueous and urea-containing solutions and antibody production, Biochem. J. 263, 365–370, 1989; Gervais, V., Guy, A., Teoule, R., and Fazakerley, G.V., Solution conformation of an oligonucleotide containing a urea deoxyribose residue in front of a thymine, Nucleic Acids Res. 20, 6455–6460, 1992; Smith, B.J., Acetic acid-urea polyacrylamide gel electrophoresis of proteins, Methods Mol. Biol. 32, 39–47, 1994; Buck, M., Radford, S.E., and Dobson, C.M., Amide hydrogen exchange in a highly denatured state. Hen egg-white lysozyme in urea, J. Mol. Biol. 237, 247–254, 1994; Shirley, B.A., Urea and guanidine hydrochloride denaturation curve, Methods Mol. Biol. 40, 177–190, 1995; Bennion, B.J. and Daggett, V., The molecular basis for the chemical denaturation of proteins by urea, Proc. Natl. Acad. Sci. USA 100, 5142–5147, 2003; Soper, A.K., Castner, E.W., and Luzar, A., Impact of urea on water structure: a clue to its properties as a denaturant? Biophys.Chem.105, 649–666, 2003; Smith, L.J., Jones, R.M., and van Gunsteren, W.F., Characterization of the denaturation of human alpha-1-lactalbumin in urea by molecule dynamics simulation, Proteins 58, 439–449, 2005; Idrissi, A., Molecular structure and dynamics of liquids: aqueous urea solutions, Spectrochim. Acta A Mol. Biomol. Spectrosc. 61, 1–17, 2005; Chow, C., Kurt, N., Murphey, R.M., and Cavagnero, S., Structural characterization of apomyoglobin self-associated species in aqueous buffer and urea solution, Biophys. J. 90, 298–309, 2006.


Lumry, R. and Rajender, S., Enthalpy-entropy compensation phenomena in water solutions of proteins and small molecules: a ubiquitous property of water, Biopolymers 9, 1125–1227, 1970; Cooke, R. and Kuntz, I.D., The properties of water in biological systems, Annu. Rev. Biophys. Bioeng. 3, 95–126, 1974; Fettiplace, R. and Haydon, D.A., Water permeability of lipid membranes, Physiol. Rev. 60, 510–550, 1980; Lewis, C.A. and Wolfenden, R., Antiproteolytic aldehydes and ketones: substituent and secondary deuterium isotope effects on equilibrium addition of water and other nucleophiles, Biochemistry 16, 4886–4890, 1977; Wolfenden, R.V., Cullis, P.M., and Southgate, C.C., Water, protein folding, and the genetic code, Science 206, 575–577, 1979; Wolfenden, R., Andersson, L., Cullis, P.M., and Southgate, C.C., Affinities of amino acid side chains for solvent water, Biochemistry 20, 849–855, 1981; Cullis, P.M. and Wolfenden, R., Affinity of nucleic acid bases for solvent water, Biochemistry 20, 3024–3028, 1981; Radzicka, A., Pedersen, L., and Wolfenden, R., Influences of solvent water on protein folding: free energies of salvation of cis and trans peptides are nearly identical, Biochemistry 27, 4538–4541, 1988; Dzingeleski, G.D. and Wolfenden, R., Hypersensitivity of an enzyme reaction to solvent water, Biochemistry 32, 9143–9147, 1993; Timasheff, S.N., The control of protein stability and association by weak interactions with water: how do solvents affect these processes? Annu. Rev. Biophys. Biomol. Struct. 22, 67–97, 1993; Wolfenden, R. and Radzcika, A., On the probability of finding a water molecule in a nonpolar cavity, Science 265, 936–937, 1994; Jayaram, B. and Jain, T., The role of water in protein–DNA recognition, Annu. Rev. Biophys. Biomol. Struct. 33, 343–361, 2004; Pace, C.N., Trevino, S., Prabhakaran, E., and Scholtz, J.M., Protein structure, stability, and solubility in water and other solvents, Philos. Trans. R. Soc. Lond. B Biol. Sci. 359, 1225–1234, 2004; Rand, R.P., Probing the role of water in protein conformation and function, Philos. Trans. R. Soc. Lond. B Biol. Sci. 359, 1277–1284, 2004; Bagchi, B., Water dynamics in the hydration layer around proteins and micelles, Chem. Rev. 105, 3179–3219, 2005; Raschke, T.M., Water structure and interactions with protein surfaces, Curr. Opin. Struct. Biol. 16, 152–159, 2006; Levy, Y. and Onuchic, J.N., Water mediation in protein folding and molecular recognition, Annu. Rev. Biophys. Biomol. Struct. 35, 389–415, 2006; Wolfenden, R., Degrees of difficulty of water-consuming reactions in the absence of enzymes, Chem. Rev. 106, 3379–3396, 2006.

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