Tetramethylbenzidine Chemical Properties |
Melting point | 168-171 °C(lit.) |
Boiling point | 100 °C |
density | 1 |
refractive index | 1.5519 (estimate) |
storage temp. | 2-8°C |
solubility | Slightly soluble. <0.1 g/100 mL at 20°C. |
pka | 4.49±0.10(Predicted) |
form | tablet |
color | Cream |
Water Solubility | Slightly soluble. <0.1 g/100 mL at 20 ºC |
Sensitive | Light Sensitive |
BRN | 2808541 |
Stability: | Stable, but moisture sensitive and may be light sensitive. Incompatible with water, strong oxidizing agents. |
CAS DataBase Reference | 54827-17-7(CAS DataBase Reference) |
EPA Substance Registry System | 3,3',5,5'-Tetramethylbenzidine (54827-17-7) |
Tetramethylbenzidine Usage And Synthesis |
Description | Tetramethylbenzidine(TMB) is a chromogenic substrate used in staining procedures in immunohistochemistry as well as being a visualising reagent used in enzyme-linked immunosorbent assays (ELISA). It is a white solid that forms a pale blue-green liquid in solution with ethyl ac etate. Tetramethylbenzidine is degraded by sunlight and by fluorescent lights. It is a soluble chromogen substrate for horseradish peroxidase detection systems. Tetramethylbenzidine is recommended for ELISA procedures. |
Chemical Properties | Tetramethylbenzidine is sensitive to prolonged exposure to light. Neutralizes acids in exothermic reactions to form salts plus water.Tetramethylbenzidine may be incompatible with isocyanates, halogenated organics, peroxides, phenols (acidic), epoxides, anhydrides, and acid halides. Flammable gaseous hydrogen may be generated in combination with strong reducing agents, such as hydrides. |
Reactions |
TMB can act as a hydrogen donor for the reduction of hydro gen peroxide to water by peroxidase enzymes such as horseradish peroxidase.
Oxidation of TMB Shows the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) to 3,3',5,5'-tetramethylbenzidine diimine The resulting diimine causes the solution to take on a blue colour, and this colour change can be read on a spectrophotometer at the wavelengths of 370 and 650 nm. |
Carcinogenicity | It is not known if Tetramethylbenzidine is carcinogenic and the evidence is contradictory: Tetramethylbenzidine is not mutagenic by the Ames test, and did not induce formation of tumors in a single-arm study of 24 rats. On that evidence, it has been used as a replacement for carcinogenic compounds such as benzidine and o-phenylenediami ne. |
storage | Keep as concentrated solution, aliquot and store at 4ºC. Do not freeze. |
Chemical Properties | White or light yellow solid |
Uses | 3,3,5,5-Tetramethyl benzidine is used as a reagent in a sensitive staining procedure for the detection of low levels of heme-associated peroxidase activity of cytochrome P-450 on SDS-polyacrylamide or agarose gel; non-carcinogenic substitute for benzidine as reagent for the detection of blood and determination of hemoglobin content. |
General Description | Pale yellow crystals or off-white powder. |
Air & Water Reactions | Insoluble in water. |
Reactivity Profile | Tetramethylbenzidine is sensitive to prolonged exposure to light . Neutralizes acids in exothermic reactions to form salts plus water. May be incompatible with isocyanates, halogenated organics, peroxides, phenols (acidic), epoxides, anhydrides, and acid halides. Flammable gaseous hydrogen may be generated in combination with strong reducing agents, such as hydrides. |
Fire Hazard | Flash point data for Tetramethylbenzidine are not available, however, Tetramethylbenzidine is probably combustible. |