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D5637 Sigma

3,3′-Diaminobenzidine tetrahydrochloride hydrate





1, 5, 10, 25, 50 g in glass bottle


DAB (3,3′-Diaminobenzidine) is utilized in many applications for visualization of peroxidase activity. In the peroxidase reaction, DAB serves as a hydrogen donor in the presence of peroxide. The oxidized DAB forms an insoluble brown end-product for use in immunohistological and immunoblotting staining procedures.

Biochem/physiol Actions

3,3′-Diaminobenzidine tetrahydrochloride hydrate has been used in the immunohistological staining procedure, western blotting and FISH (fluorescence in situ hybridization).

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Hazard statements 
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NONH for all modes of transport
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Frequently Asked Questions

What is Product D5637, 3,3′-Diaminobenzidine tetrahydrochloride hydrate (DAB), soluble in?
DAB HCl is soluble in water at 50 mg/ml yielding a clear to slightly hazy solution. For consistency, it is important to allow DAB to come to room temperature before making solutions. Water quality is extremely important, so only high quality deionized or distilled water should be used to make DAB solutions. Please click on the following link to view product information sheet on our website: http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Product_Information_Sheet/d5637pis.Par.0001.File.tmp/d5637pis.pdf
What is the solution stability of Product D5637, 3,3′-Diaminobenzidine tetrahydrochloride hydrate (DAB)?
In our experience, solutions of DAB should be made immediately prior to use for best results. If and only if necessary, solutions can be kept in the dark and refrigerated for overnight storage. Based on our Immunology Department’s experience, storing the solution overnight did not give good reproducibility. However, the literature indicates that it is possible to store solutions of DAB at -20°C. DAB dissolved in Tris buffered saline (pH 7.6) at 0.6 mg/ml was reported to be stable for several weeks at 4°C. This information is available in our product information sheet, which one can view by clicking on the following link: http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Product_Information_Sheet/d5637pis.Par.0001.File.tmp/d5637pis.pdf
Can the solutions of Product D5637, 3,3′-Diaminobenzidine tetrahydrochloride hydrate (DAB), be filtered?
DAB solution may be filtered through a 0.2 micron filter immediately prior to use.
Do you have a staining protocol for Product D5637, 3,3′-Diaminobenzidine tetrahydrochloride hydrate (DAB)?
Dissolve 10 mg in 15 mL of Tris buffered saline, pH 7.6 (Tris-buffered saline tablet, Sigma Product No. T5030 may be used). Add 12 uL of fresh 30% hydrogen peroxide (Sigma Product No. H1009) prior to use. Solution may be filtered through a 0.2 micron filter immediately prior to use if necessary. DAB and hydrogen peroxide concentrations may be adjusted to suit individual applications. The stain can be enhanced with Ni. Make up 0.3% (w/v) NiCl2 in water. Add 1 mL of the NiCl2 stock to 9 mL DAB solution.
What is the shelf life of Product D5637, 3,3′-Diaminobenzidine tetrahydrochloride hydrate (DAB)?
This product has a shelf life of 3 years. Click on the following link to view this information on the specification sheet: http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Product_Information_Sheet/d5637pis.Par.0001.File.tmp/d5637pis.pdf
How is Product D5637, 3,3′-Diaminobenzidine tetrahydrochloride hydrate (DAB), shipped and stored?
The product is shipped at ambient temperature. However, it is stored at -20 degrees C for long term storage.
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
How do I find price and availability?
There are several ways to find pricing and availability for our products.  Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers. 
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS. To access the shipping information for this material, use the link on the product detail page for the product, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
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Keywords: Applications, Enzyme-linked immunosorbent assay, Formulations, Immunohistochemistry, Western blot

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