Enhanced DAB Substrate Kit

The Kit contains the following reagents to make an active stable substrate solution for use with the enzyme horseradish peroxidase:

1. 6 ml Buffer Stock Solution (Storage 4°C)
2. 12 ml Solutle DAB Stock Solution (Storage -20°C)
3. 6 ml Hydrogen Peroxide Stock Solution (Storage 4°C)
4. 6 ml Enhancing Stock solution (Storage 4°C)

Store DAB Stock Solution below -20°C. The remaining reagents are stored at 4°C

Next Day 2-8°C

DAB Substrate (3,3/-Diaminobenzidine) is the most commonly used substrate for demonstrating the presence of horseradish peroxidase in immunohistochemistry or immunoblotting. It produces a stable water/alcohol insoluble reddish brown product. This effect is enhanced by modifying the DAB solution with the addition of metal ions to produce a more dense reaction forming a localized deep blue/black precipitate.1,2 Leinco's Metal enhanced DAB Substrate Kit is a stable solution developed to give enhanced color reaction with low background. The reagents supplied are sufficient for making 300ml working solution.

Warning: As DAB and nickel are suspected carcinogens care should be taken in handling and disposal of the working solution. Gloves, lab coats and good laboratory procedures are necessary when using the reagents. For disposal the working solution is discarded in 3% potassium permanganate (KMNO₄) and 2% sodium carbonate in distilled water. Wash out all containers used for substrate staining with this solution. Dispose according to local regulations.

1. (1981) J. Histochem. Cytochem. Vol.29, No.6:775 2. (1989) J. Clin, Pathol, 42:875-880