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3,3,5,5-Tetramethylbenzidine or TMB is a chromogenic substrate used in staining procedures in immunohistochemistry as well as being a visualising reagent used in enzyme-linked immunosorbent assays. 3,3?,5,5?-Tetramethylbenzidine (TMB) is a noncarcinogenic substitute (Ames test negative) for benzidine suitable as peroxidase substrate for use in ELISA procedures. The substrate produces a soluble end product that is pale blue in color and can be read spectrophotometrically at 370 or 620-650 nm. The TMB reaction may be stopped with 2 M H2SO4 (resulting in a yellow color), and read at 450 nm. A sensitive and specific reagent for the detection of blood assay of hemoglobin assay of peroxidases.
J C Standefer,; D Vanderjagt. Use of tetramethylbenzidine in plasma hemoglobin assay.. Clinical Chemistry. 1977, 23 (4), 749-751.
R C Lijana,; M C Williams. Tetramethylbenzidine--a substitute for benzidine in hemoglobin analysis.. Journal of Laboratory and Clinical Medicine. 1979, 94 (2), 266-276.