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3,3'-Diaminobenzidine tetrahydrochloride, 1 g, glass

≥98 %, p.a.
Examples of effect: Lead to damage to heath, cause irritation to eyes, skin or respiratory organs. Are fatal in larger quantities. Safety: As previously stated, in the event of skin irritation or contact with eyes, rinse with water or a suitable medium.
Examples of effect: Have an allergenic effect, carcinogenic, mutagenic, toxic to reproduction and cause developmental toxicity (reprotoxic) or damage to organs Safety: You must be well-informed before starting work with these substances; wear protective clothing and gloves, eye protection and mask or breathing protection.
Warning
H315-H319-H335-H341-H351
i causes skin irritation, causes serious eye irritation, may cause respiratory irritation, suspected of causing genetic defects, suspected of causing cancer
P202 P261 P280 P305+P351+P338 P308+P313
i do not handle until all safety precautions have been read and understood, avoid breathing dust, wear protective gloves/eye protection, IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing, IF exposed or concerned: Get medical advice/attention
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DAB, 3,3',4,4'-Tetra-aminobiphenyl tetrahydrochloride
Empirical formula C12H14N4 · 4 HCl
Molar mass (M) 360,10 g/mol
Melting point (mp) >280 °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.
Application: Stock solution: 1-2 % (10-20 mg/ml) in 50 mM Tris buffer, pH 7.3. Storage of stock solution in aliquots at -20 °C. 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.

Popular substrate of peroxidases. DAB is used for biochemical and histological detection of endogeno ...
Product details
Type analysis

€24.65/Pack Qty. 

excl. VAT. | 1 g per Pack Qty.

Art. No. CN75.1

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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.



Technical Information
Application Western, in situ, Histochemistry  
Enzyme Meerrettich Peroxidase  
Detection Farbe (braun)  
Signal product Präzipitat (Blot)  
3,3'-Diaminobenzidine tetrahydrochloride
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CN75.1 1 g glass €24.65
CN75.2 5 g plastic €79.45
CN75.3 10 g plastic €136.00
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General information

Assay protocol for use of BCIP/NBT in immunoblot procedures:

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.


Detection of peroxidase activity (Immunoassay):

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.


Analysis certificates

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The following analysis certificates have been found:

Type analysis

Assay (HPLC)≥98 %  
Suitablility for determination of Seleniumcomplies  
Appearance of solutionReddish brown  

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