Application

3,3′,5,5′-Tetramethylbenzidine (TMB) is a peroxidase substrate suitable 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.

3,3′,5,5′-Tetramethylbenzidine dihydrochloride has been used as a substrate for theanti-mouse IgG (? specific) horseradish peroxidase-conjugated antibody in the ELISA of mice seraanti-mouse peroxidase antibody in the protein tyrosine phosphatase assay of naive and effector T cell lysatein enzymatic hydroperoxide (EH) assay and in prooxidants–antioxidants balance (PAB) assay of serum samples from diabetic patients