05 Cell Staining
10 Redox Dyes

DAB

DAB

Coloring Substrate for Peroxidase

  • Product code
    D006  DAB
  • CAS No.
    7411-49-6
  • Chemical name
    3,3'-Diaminobenzidine, tetrahydrOchloride
  • MW
    C12H18Cl4N4=360.11
Unit size Price Item Code
1 g $234.00 D006-10
5 g $859.00 D006-12
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Product Description

DAB is one of the most commonly used oxidative chromogenic dyes for the detection of peroxidase in immunohistochemistry. In the presence of H2O2 and peroxidase, DAB is oxidized into a brown pigment. These brown pigments are aniline black type compounds and are firmly deposited around peroxidase on cell membranes or tissues. Most DAB available on the market appears brown as a result of oxidation during processing or storage. On the other hand, Dojindo offers a high quality DAB with a white or slightly reddish-gray powder appearance that is good for sharp staining.

Technical info

Preparation of Sample Staining Solution

1. Dissolve 9 mg DAB with 1 ml PBS to prepare 100X DAB solution.
2. Mix 5 μl of 30% hydrogen peroxidase solution with 1 ml PBS to prepare 200X H2O2 solution.
3. Add 10 ul of 100X DAB solution and 5 ul of 200X H2O2 solution to 1 ml PBS to prepare staining solutiona).
4. Add the staining solution to a sample in a staining chamber, and incubate the sample at room temperature for 5 minutes to 1 hourb).
5. Wash the sample with PBS several times to stop the staining reaction.

a)The staining solution is not stable. Prepare fresh solution prior to use.
b)For better staining, incubate the sample with 0.09 mg DAB/PBS solution for 10 minutes before adding the staining solution.

References

Open References

1. A. B. Novikoff, et al., Studies on Microperoxisomes V. Are Microperoxisomes Ubiquitous in Mammalian Cells. J Histochem Cytochem. 1973;21:737.
2. V. Herzog, et al., A New Sensitive Colorimetric Assay for Peroxidaase Using 3, 3 EDiaminobenzidine as Hydrogen Donor. Anal Biochem. 1973;55:554.
3. H. Yamada, et al., Improvement of Technique of Immunohistochemical Demonstraction of Bioactive Substances in the Central Nervous System. Acta Histochem Cytochem. 1987;20:629.
4. K. L. Cheng, Determination of Traces of Selenium 3,3 EDiaminobenzidine as Selenium(IV) Organic Reagent. Anal Chem. 1956;28:1738.

Handling and storage condition

Specification
Appearance: White to pinkish gray powder
Purity (Titration): ≧ 97.0 % (as dry substance)
Solubility in water: To pass test
Solubility in Tris buffer: To pass test
Absorbance (Se complex): ≧ 0.500 (around 420 nm)
Loss on drying: ≦ 5.0 %
Sulfated ash: ≦ 0.05 %
IR spectrum: Authentic
Handling and storage condition
0-5°C, Protect from light
Danger / harmful
symbol mark
健康有害性
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