Technical Data Sheet
3,3',5,5'-Tetramethylbenzidine dihydrochloride, 500 mg
Molar mass (M) 313,27 g/mol
Melting point (mp) 300 °C
Storage temp. -20 °C
Transport temp. ambient temp.
WGK 1
CAS No. 207738-08-7
EG-Nr. 264-769-6
Storage of stock sol.: At +4 °C if tightly sealed. Oxidation can be seen by colour-shift to a faint blue.
Water-soluble peroxidase substrate
€53.65/Pack Qty.
excl. VAT. | 500 mg per Pack Qty.
Art. No. 4107.1
Product details
3,3',5,5'-Tetramethylbenzidine dihydrochloride ≥99 %, for biochemistry
Tetramethylbenzidine dihydrochloride is used as highly sensitive substrate for detection of horseradish peroxidase labelled probes, particularly in solution-based assays like ELISA techniques. It produces a soluble, blue-coloured end product, which can be read spectrophotometrically.
The dihydrochloride form is well water-soluble, making it a versatile tool in laborartory science.
Working solution: Immediately prior to use add 2 µl / 10 ml of 30 % hydrogen peroxide (Liem et al. (1979) Anal Biochem, 98:388-93).
| Enzyme | Horseradish peroxidase |
| Detection | Colour (blue) |
| Signal product | soluble (ELISA) |
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| Art. No. | Pack Qty. | Pack. | Price | Quantity | |
|---|---|---|---|---|---|
| 4107.1 | 500 mg | glass |
€53.65 |
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| 4107.2 | 1 g | glass |
€91.30 |
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In stock
Available (note delivery time)
In procurement
No longer available
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- Subtotal: 0.00
Downloads / MSDS
General information
Stock solutions (All solutions are stable for at least 1 year at 4 °C):
0.5 g NBT in 10 ml 70 % dimethylformamide;
0.5 g BCIP (p-toluidine salt) in 10 ml 100 % dimethylformamide.
Incubation buffer for alkaline phosphatase:
100 mM NaCl, 5 mM MgCl2,100 mM Tris (pH 9.5).
Fresh substrate solution: 66 μl NBT stock solution + 10 ml incubation buffer, mix well, add 33 μl BCIP stock solution. Use within 1 hour.
Blot development: Approx. 10 ml substrate solution per 15 x 15 cm2 membrane surface. Develop at room temperature until bands become visible (approx. 30 mins).
Reaction stop: Rinse with PBS/20 mM EDTA.
Dissolve 1 mg TMB in 0.1 ml dimethylsulfoxide (Art. No. 4720); add 9.9 ml of a 0.1 M sodium acetate solution (pH 6.0) (Powder: Art. No. 6779), filter and add H2O2 (Art. No. 8070) (final concentration 0.01 %).
Always prepare freshly!
Incubation 10-30 mins at room temperature (approx. 50 μl per microtitre well; finally add 50 μl 1 M H2SO4, Art. No. X873, per well).
Photometric quantitation at 450 nm.
Reference: Bos E.S. et al., 1981, J. Immunoassay 2, (3/4), 187.
Certificates of Analysis
Guarantee analysis
| Assay (titr.) | ≥99 % |
| Solution (1 % in methanol) | colourless to yellow-green |
| Water (KF) | ≤5 % |
| Sulphated ash | ≤0.1 % |