Rev. 9/30/97
The DAB reaction is performed to deposit a brown chromagen substrate where there are peroxide enzymes present.
DAB is a hazardous substance. All items in contact with the solution must be thoroughly washed or disposed of in the DAB dry waste container stored in the hood. DAB solutions are to be disposed of in the DAB liquid wasted container stored in the hood.
Obtain a beaker of sufficient size and measure a quantity of .1M Tris buffer to cover slides and or tissue
Prepare a suitable quantity of DAB solution by adding 1 tube of prediluted DAB (12.5 mg/250µl) per 50 ml into the .1 M Tris buffer.
Allow solution to mix well using a stir bar.
Place slides and or tissue into solution and allow to incubate for about 5 minutes.
Add 6 µls of 30% H2O2 (or 60 µls of 3% H2O2) per 100 mls of solution and mix well.
Allow DAB reaction to occur for 5-15 minutes. Slides and tissue can be visualized to confirm reaction product is present.
Following the reaction wash slides and tissue with at least 3 changes of buffer.
The substrate is resistant to both aqueous and organic solvents.