The Use of 3, 3′, 5, 5′-Tetramethylbenzidine as a Peroxidase Substrate in Microplate Enzyme-Linked Immunosorbent Assay

Abstract
The assay conditions for the use of 3)3′,5,5′ tetramethylbenzidine (TMB) in microplate enzyme-linked immunosorbent assay are described. TMB is a safe (non-mutagenic) chromogen that is more sensitive than OPD as a substrate for horseradish peroxidase. We describe the optimum storage and assay conditions for this chromogen.