3,3′,5,5′-Tetramethylbenzidine peroxidase substrate, chromogenic, liquid

Synonym(s): TMB

General description

3,3′,5,5′-Tetramethylbenzidine (TMB), Liquid Peroxidase SubstrateA high-performance chromogenic substrate designed for ELISA systems using horseradish peroxidase (HRP) conjugates. This ready-to-use liquid substrate generates a soluble blue reaction product upon oxidation by HRP, enabling sensitive detection in microwell assays. At assay endpoint, addition of acid converts the blue product to a stable yellow chromophore measurable at 450 nm, allowing precise quantification of target analytes.

This TMB substrate is light-sensitive, high-purity, and ideal for diagnostic, research, and analytical workflows requiring reliable HRP-based signal development. Application TMB substrate is used in ELISA procedures, which utilize horseradish peroxidase conjugates. TMB substrate develops a soluble blue reaction product that can be read at 370 or 655 nm at 370 or 655 nm. The reaction is stopped with acid, forming a yellow reaction product which enables accurate intensity measurement at 450 nm.

How to use our TMB substrate:

Our TMB substrate is supplied as a ready-to-use peroxidase substrate containing TMB in a mildly acidic buffer. Prior to reaction with peroxidase, the substrate should be a colorless to light bluish-green solution. The substrate system develops a blue reaction product when reacted with peroxidase. 

Upon reaching your assay endpoint with addition of acid, measure the TMB substrate intensity at 450 nm.

Our TMB Substrate is recommended for ELISA (microwell) procedures, and has been used in various assays, as a substrate for horseradish peroxidase antibody conjugate for detecting hendra virus glycoprotein, tetracosactide, sIgA and neutrophil extracellular traps (NETs).

3,3′,5,5′-Tetramethylbenzidine (TMB) Liquid Substrate System is not recommended for membrane applications. Since TMB substrate produces a soluble reaction product, it is not recommended for histochemistry or blotting.

Note:TMB substrate is light-sensitive and should be protected from direct sunlight or UV sources.