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TMB dihydrochloride(Chromogenic Substrate) CS120 3,3',5,5'-Tetramethylbenzidene dihydrochloride[ 64285-73-0 ] f.w.: 349.3 daltons |
3,3',5,5'-Tetramethylbenzidine dihydrochloride dihydrate (TMB) is an aromatic amine that undergoes oxidation by the higher oxidation states of heme peroxidases thereby serving as a reducing co-substrate. One electron oxidation of TMB results in a radical cation that creates a charge-transfer complex with the unoxidized compound. This charge transfer complex absorbs at 652 nm (? = 39,000). The completely oxidized form (diimine) absorbs at 450 nm (? = 59,000) and is formed by two sequential one-electron oxidations of TMB. Thus the stoichiometry of oxidation is 0.5 mole charge transfer complex (?max = 652 nm) or 1 mole of diimine (?max = 450 nm) formed (or TMB oxidized) per mole of hydroperoxide reduced by the peroxidase.
1 Josephy, P.D., Eling, T., Mason, R.P. The horseradish peroxidase-catalyzed oxidation of 3,5,3',5'-tetramethylbenzidine. Free radical and charge-transfer complex intermediates. J Biol Chem 257 3669-3675 (1982). 2 Marquez, L.A., Dunford, H.B. Mechanism of the oxidation of 3,5,3',5'-tetramethylbenzidine by myeloperxidase determined by transient-and steady-state kinetics. Biochemistry 36 9349-9355 (1997).