TMB Substrate (Single-Component)

TMB Substrate is a single-step and single-component ready-to-use substrate solution containing soluble Tetramethylbenzidine (TMB). TMB is a chromogenic substrate used in ELISA, IHC & Western Blot applications to detect the antibodies/proteins conjugated with horseradish peroxidase (HRP) enzyme. Once oxidized by the enzyme, TMB substrate yields a blue product that absorbs at 370nm and 652nm. Adding sulfuric or phosphoric acid to stop the reaction changes the product to yellow that absorbs at 450nm. TMB Substrate produces a very large signal-to-noise ratio, although its high sensitivity requires careful washing and blocking to avoid background staining. When performing IHC techniques, aqueous counter stains and mounting medium must be used.