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DAB Substrate

dark brown, /black precipitate; Visually evaluated

3,3′-diaminobenzidine tetrahydrochloride, DAB



Quality Level


dark brown, /black precipitate; Visually evaluated

storage temp.


General description

The metal enhanced DAB Substrate utilizes cobalt chloride and nickel chloride in a special formulation to produce a dark brown/black precipitate in the presence of horseradish peroxidase (HRP).
3, 3′ diaminobenzidine tetrahydrochloride (DAB) is a carcinogen that should be used safely. It is one of the major sensitive substrates for HRP (horseradish peroxidase). DAB is brown or amber in color and becomes insoluble and precipitates over the antigen-antibody site due to the presence of hydrogen peroxide and peroxidase.


  • DAB Substrate is a color sustrate for peroxidases and is used in a variety of applications:Immunohistocytochemistry
  • In situ hybridization
  • Western blot
  • Dot blot
DAB substrate has been used in immunofluorescence and western blot.


1 set containing 2 components.

Preparation Note

Working solution:
  1. Determine the volume of substrate required for the development of peroxidase.
  2. Remove the DAB/metal concentrate (10x) from -15 to -25 °C storage.
    Note: Do not bring to 15 to 25 °C.
  3. Dilute the DAB/metal concentrate (10x) with the peroxide buffer to a 1x working solution. For example, if 5 ml of substrate are required, dilute 500 μl of the DAB/metal concentrate (10x) with 4.5 ml of the Peroxide buffer.
  4. Mix well.
  5. Add the 1x working solution to the tissue or blot until desired substrate development.
    Note: The 1x solution is stable over many hours.
    For best results, store the working solution at 2 to 8 °C when not using.

Storage conditions (working solution): 2 to 8 °C
For best results, store the working solution at 2 to 8 °C when not using.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Kit Components Only

Product No.

  • DAB/metal concentrate, 25 ml 10x concentrated

  • Peroxide Buffer, 250 ml

Signal Word


Hazard Classifications

Acute Tox. 3 Dermal - Acute Tox. 3 Inhalation - Acute Tox. 4 - Aquatic Chronic 2 - Carc. 1A - Carc. 1B - Flam. Liq. 2 Oral - Repr. 1B - Skin Sens. 1 Inhalation - STOT RE 2 - STOT SE 1

Storage Class Code

3 - Flammable liquids



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