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Safety Information

D5637

Sigma-Aldrich

3,3′-Diaminobenzidine tetrahydrochloride hydrate

≥96%

Synonym(s):

(1,1′-Biphenyl)-3,3′,4,4′-tetramine tetrahydrochloride dihydrate

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About This Item

Linear Formula:
C12H14N4 · 4HCl · xH2O
CAS Number:
EC Number:
MDL number:
UNSPSC Code:
12171500
PubChem Substance ID:
NACRES:
NA.47

Quality Level

Assay

≥96%

form

powder, crystals or chunks

technique(s)

titration: 96-104% (with NaOH)

color

white to light brown, and Faint Red and Faint Grey to Light Grey

mp

280 °C

solubility

H2O: 50 mg/mL

application(s)

diagnostic assay manufacturing
hematology
histology

storage temp.

−20°C

SMILES string

O.Cl.Cl.Cl.Cl.Nc1ccc(cc1N)-c2ccc(N)c(N)c2

InChI

1S/C12H14N4.4ClH.H2O/c13-9-3-1-7(5-11(9)15)8-2-4-10(14)12(16)6-8;;;;;/h1-6H,13-16H2;4*1H;1H2

InChI key

DXWSCXIZIHILNP-UHFFFAOYSA-N

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General description

3,3′-Diaminobenzidine tetrahydrochloride hydrate is a chromogen commonly used in immunohistochemistry and other staining techniques to visualize the presence of antigens or antibodies in biological samples. DAB HCl is designed as an immunohistology stain to visualize peroxidase activity in cells. DAB oxidation can be measured using a spectrophotometer.

Application

3,3′-Diaminobenzidine tetrahydrochloride hydrate is suitable for immunohistology staining procedures, western blotting, and FISH (fluorescence in situ hybridization). DAB is used in studying epithelial cell apoptosis in the event of early cataracts, and immunohistochemistry protocols of autosomal dominant polycystic kidney disease (PKD).

Biochem/physiol Actions

In the peroxidase reaction, DAB serves as a hydrogen donor in the presence of peroxide. The oxidized DAB forms an insoluble brown-colored complex.

Pictograms

Health hazardExclamation mark

Signal Word

Danger

Hazard Statements

Hazard Classifications

Acute Tox. 4 Oral - Carc. 1B - Eye Irrit. 2 - Muta. 2

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Regulatory Information

危险化学品

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. What is the shelf life of Product D5637, 3,3´-Diaminobenzidine tetrahydrochloride hydrate (DAB)?

    This product has a shelf life of 3 years.

  6. 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.

  7. 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.

  8. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  9. 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.

  10. 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. 

  11. My question is not addressed here, how can I contact Technical Service for assistance?

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A new sensitive colorimetric assay for peroxidase using 3,3'-diaminobenzidine as hydrogen donor.
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Mammalian AMP-activated protein kinase subfamily.
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NBT-BCIP substrate system aids in western blotting and immunohistological staining, producing a blue-purple insoluble end product.

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