Excellent colorimetric substrate for detection of horseradish peroxidase labelled probes; particularly useful in ELISA techniques, blue result can be read spectrophotometrically at 370 or 650 nm3,3',5,5'-Tetramethylbenzidine is used as a clinical reagent for testing blood. It is also a chromogenic substrate which is used in staining procedures in immunohistochemistry. Further, it is used as a visualizing reagent in enzyme-linked immunosorbent assays. In addition to this, it is used as a replacement for carcinogenic compounds such as benzidine and o-phenylenediamine.
This Thermo Scientific Chemicals brand product was originally part of the Alfa Aesar product portfolio. Some documentation and label information may refer to the legacy brand. The original Alfa Aesar product / item code or SKU reference has not changed as a part of the brand transition to Thermo Scientific Chemicals.
General Description
• 3,3',5,5'-Tetramethylbenzidine, also known as 4,4'-Diamino-3,3',5,5'-tetramethylbiphenyl or TMB, is a colorimetric substrate for detection of horseradish peroxidase-labeled probes
• Oxidation of 3,3',5,5'-Tetramethylbenzidine results in a pale blue end product that can be detected at 370 or 655 nm. Sulfuric acid (H2SO4) is used to stop the oxidation reaction and stabilize the reading, resulting in a yellow color that can be read at 450 nm
Application
• 3,3',5,5'-Tetramethylbenzidine is used as a chromogenic visualization agent in enzyme-linked immunosorbent assays (ELISA) or immunohistochemistry (IHC) techniques
• 3,3',5,5'-Tetramethylbenzidine has been used to detect molecules in aqueous samples such as copper ion and hypochlorite
RUO – Research Use Only
Literature References
Tang, S.; Chang, Y.; Chia, G. H.; Lee, H. K. Selective extraction and release using (EDTA-Ni)-layered double hydroxide coupled with catalytic oxidation of 3,3',5,5'-tetramethylbenzidine for sensitive detection of copper ion. Anal. Chim. Acta 2015, 885, 106-113.
Guo, Y.; Ma, Q.; Cao, F.; Zhao, Q.; Ji, X. Colorimetric detection of hypochlorite in tap water based on the oxidation of 3,3',5,5'-tetramethylbenzidine. Anal. Methods 2015, 7 (10), 4055-4058.
Frey, A.; Meckelein, B.; Externest, D.; Schmidt, M.A. A stable and highly sensitive 3,3',5,5'-tetramethylbenzidine-based substrate reagent for enzyme-linked immunosorbent assays. J Immunol Methods. 2000, 233(1-2):47-56.