• USA Home
  • T8665 - 3,3′,5,5′-Tetramethylbenzidine (TMB) Liquid Substrate System

T8665 Sigma

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

peroxidase substrate

Synonym: TMB




This substrate is very light-sensitive and should be protected from direct sunlight or UV sources.

Physical form



TMB is a substrate for horseradish peroxidase. Forms a soluble blue end product that may be read at 370 nm or 655 nm. The TMB reaction may be stopped with 0.5 M H2SO4, resulting in a yellow color, and read at 450 nm. Recommended for ELISA (microwell) procedures, not recommended for membrane applications.
3,3′,5,5′-Tetramethylbenzidine has been used in a study to demonstrate that by measuring substrate absorption values off the absorption maximum, the measuring range of any immunoenzymometric assay can be extended significantly without jeopardizing assay characteristics. 3,3′,5,5′-Tetramethylbenzidine has also been used in a study to understand the formation of CuS concave superstructures with peroxidase-like activity.

Price and Availability

Amplified Detection
Safety & Documentation

Safety Information

Signal word 
Hazard statements 
Precautionary statements 
UN 1993C 3 / PGIII
WGK Germany 
Flash Point(F) 
129.2 °F
Flash Point(C) 
54 °C
Protocols & Articles


Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Colorimetric Alkaline Phosphatase and Peroxidase Substrate Detection Systems

Nitroblue Tetrazolium (NBT) is used with the alkaline phosphatase substrate 5-Bromo- 4-Chloro-3-Indolyl Phosphate (BCIP) in western blotting and immunohistological staining procedures. These substrat...
BioFiles 2008, 3.4, 6.
Keywords: Applications, Enzyme-linked immunosorbent assay, Formulations, Immunohistochemistry, Western blot

Related Content

Antibody Explorer - Over 80,000 Products

Monoclonal and polyclonal primary antibodies are focused on cell biology, neurobiology and molecular biology. Secondary antibodies targeting multiple host’s IgG are conjugated to alkaline phosphatase...
Keywords: Amplification, Buffers, Cell biology, Enzyme-linked immunosorbent assay, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Molecular biology, Phosphorylations, Purification, Western blot

Peer-Reviewed Papers


Set your institution to view full text papers.
Related Products

Technical Service:

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Bulk Ordering & Pricing:

Need larger quantities for your development, manufacturing or research applications?