Technical Data Sheet
3,3'-Diaminobenzidine tetrahydrochloride, 10 g, plastic
Molar mass (M) 360,10 g/mol
Boiling point (bp) 481,7 °C
Flash point (flp) 282,7 °C
Melting point (mp) 300 °C
Storage temp. +4 °C
WGK 1
CAS No. 7411-49-6
EG-Nr. 231-018-9
Substrate of horseradish peroxidase. For detection of endogenous peroxidase and for immunohistochemical stainings.
DAB is oxidised by HRP, forming a brown precipitate, which is unsoluble in aqueous and organic solvents. This precipitate can be detected in visible light and does not bleach during long-term storage.
€177.40/Pack Qty.
excl. VAT. | 10 g per Pack Qty.
Art. No. CN75.3
Alternative products
Product details
3,3'-Diaminobenzidine tetrahydrochloride ≥98 %, p.a.
Popular substrate of peroxidases. DAB is used for biochemical and histological detection of endogenous peroxidases like catalase and cytochome oxidase. Additionally, it is the most common substrate for horseradish peroxidase (HRP) in histochemical and cytological assays.
DAB is oxidised by HRP, forming a brown precipitate, which is unsoluble in aqueous and organic solvents. This precipitate can be detected in visible light and does not bleach during long-term storage.
Working concentration: 0.05-0.1 % (0.5-1 mg/ml) DAB in buffer (PBS, TBS, pH 7.0-7.6) containing 0.01 % hydrogen peroxide. Always use freshly prepared working solution. For oxidation high concentrations of fresh hydrogen peroxide are needed.
| Application | Western, in situ, Histochemistry |
| Enzyme | Horseradish Peroxidase |
| Detection | Colour (brown) |
| Signal product | Precipitate (Blot) |
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| Art. No. | Pack Qty. | Pack. | Price | Quantity | |
|---|---|---|---|---|---|
| CN75.1 | 1 g | glass |
€31.10 |
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| CN75.2 | 5 g | plastic |
€100.55 |
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| CN75.3 | 10 g | plastic |
€177.40 |
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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
| Assay | ≥98 % |
| Suitablility for determination of Selenium | complies |
| Appearance | off-white or brow powder |