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3,3′,5,5′-Tetramethylbenzidine (TMB) Liquid Substrate System for ELISA

peroxidase substrate

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General description

TMB substrate is a chromogenic substrate for horseradish peroxidase - HRP.


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.


Storage Class Code

10 - Combustible liquids



Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

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