TMB (3,3 ,5,5 -Tetramethylbenzidine) is a widely used chromogen for HRP-based detection systems, such as WB and ELISA. After its reaction with peroxidase, a soluble blue reaction product is obtained.

Store in dark at 4 ℃.

TMB substrate solution preparation for ELISA Buffer A solution preparation: 205 mM potassium citrate, pH 4.0 3.075 mM H₂O₂ Note: The mixture of two components of Buffer A should be freshly done before each experiment and be sterile-filtered (pore size 0.22μm). Solution B stock solution preparation: 41 mM TMB (CSN12373) 8.2 mM TBABH (CAS 33725-74-5) Dissolve in DMA (CSN23648) Solution B can be stored in dark at 4°C up to 16 months. Working solution: Add 200 μL Solution B to 8 ml Buffer A 10-20 min before use. The detection limit of this working solution is 24.2 pg/mL protein (defined as the endpoint of the anti-ovalbumin IgG dilution series) Absorbance value should be read at 450 nm.{{Frey A, Meckelein B, Externest D, Schmidt MA. A stable and highly sensitive 3,3',5,5'-tetramethylbenzidine-based substrate reagent for enzyme-linked immunosorbent assays. J Immunol Methods. 2000 Jan 13;233(1-2):47-56. doi: 10.1016/s0022-1759(99)00166-0. PMID: 10648855.|https://pub

TMB (3,3 ,5,5 -Tetramethylbenzidine) is a widely used chromogen for HRP-based detection systems, such as WB and ELISA. After its reaction with peroxidase, a soluble blue reaction product is obtained.