| ImmunoTek(TM) TMB (3,3,5,5-Tetramethylbenzidine) Stable Liquid Substrate for HRPO, 1.25mM, Soluble Tetramethylbenzidine (TMB) is an excellent substrate for detecting horseradish peroxidase (HRP) labeled probes. TMB produces a soluble end product which is blue in color that is quantitated at 650nm, providing an ultrasensitive quantitative substrate system. TMB has been shown to be a safe-sensitive substrate for the assay of horseradish peroxidase (HRP). Initially, in the presence of HRP and hydrogen peroxide, a one-electron oxidation product is formed. This compound, a cation free radical, is blue in color wth an adsorption maximum at 653nm. Further reaction with HRP/H2O2 or acidification of the radical with acid yields the diimine terminal oxidation product adsorbing light at 450 nm. The extinction coefficient of the radical (E653 nm = 5.9x10e4 mol-1 cm-1) provide a remarkably sensitive system for the assay of HRP and HRP labeled probes. TMB is a single component reagent stable at room temperature and not sensitive to normal laboratory light. It is optimized with respect to TMB and hydrogen peroxide concentrations and yields a linear response with the concentrations of HRP usually employed in immunologic assays. Application: Kinetic and Endpoint Assays of Horseradish Peroxidase Method: After reacting with a HRP labeled probe, tissue sections or blots are incubated in the TMB solution. A one electron oxidation occurs resulting in the formation of a stable blue precipitate at the sites of HRP activity. Features: Single component system Room temperature stable Exhibits low background levels Safe to use - no weighing of hazardous powders Save preparation time - use right out of the bottle Ultra-low sensitivity Minimal lot-to-lot variation Reagent Provided: Tetramethylbenzidine Solution: Contains TMB, 1.25mM/L in an acetate buffer with proprietary stabilizers and precipitating agents. Storage and Stability: Stable for 12 months at RT. To extend the stability of product, refrigerate at 4C. |
