Technical Data Sheet
3,3',5,5'-Tetramethylbenzidine, 1 g, glass
Molar mass (M) 240,35 g/mol
Melting point (mp) 170 °C
Storage temp. +4 °C
WGK 1
CAS No. 54827-17-7
EG-Nr. 259-364-6
Substrate for peroxidase
€65.05/Pack Qty.
excl. VAT. | 1 g per Pack Qty.
Art. No. 6350.1
Product details
3,3',5,5'-Tetramethylbenzidine ≥98 %, p.a.
Used as a highly sensitive substrate for peroxidases, particularly in ELISA procedures. Can replace carcinogenic benzidine.
Dissolves easily in acetone, chloroform and ethanol.
Working solution: 1 mg TMB in 0.1 ml dimethyl sulfoxide, add 9.9 ml 0.1 M sodium acetate (pH 6). Filter and add H2O2 ad 0.01 %. Always prepare freshly!
Application: approx. 50 µl per 96well, incubate for 10-30 mins. at room temperature. Add 50 µl 1 M H2SO4 and quantitate photometrically at 450 nm (acc. to: Bos et. al. (1981) J. Immunoassay 2:187).
Protect from light.
| Enzyme | Horseradish Peroxidase |
| Detection | Colour (blue) |
| Signal product | soluble (ELISA) |
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| Art. No. | Pack Qty. | Pack. | Price | Quantity | |
|---|---|---|---|---|---|
| 6350.1 | 1 g | glass |
€65.05 |
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| 6350.2 | 5 g | plastic |
€198.90 |
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| 6350.3 | 25 g | plastic |
€665.45 |
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In stock
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In procurement
No longer available
Delivery date currently unknown
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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
Type analysis
| Appearance | beige cryst. powder |
| Assay (GC) | ≥99.0 % |
| Sulphated ash | ≤0,2 % |
| Loss on drying (3 h, 60 °C, vacuum) | ≤1,0 % |
| Melting range | 166-171 °C |
| Can replace carcinogenic benzidine. | |