Medico DAB Substrate (3, 3’-diaminobenzidine) produces a brown reaction product in the presence of peroxidase (HRP) enzyme. Substrate Kit contains all of the reagents necessary toprepare. These reagents are supplied in convenient dropper bottles. REAGENTS
Content Size Storage Reagents A DAB 20×concentrated solution 3ml -20℃ Reagents B H2O2 20×concentrated solution 3ml 4-8℃ Reagents C TBS 20× concentrated solution 3ml 4-8℃ STORAGE
- Store reagents in original bottles at the required conditions (one year)
- Avoid storing reagents or working solution in strong direct light.
PREPARATION OF SUBSTRATE WORKING SOLUTION
- To 1.0 ml of distilled water
- Add 50 µl of Reagents C
- Add 50 µl of Reagents A
- Add 50 µl of Reagents B
- Mix well before use
INSTRUCTIONS FOR USE For Tissues or Cells After incubation with a peroxidase (HRP) detection system, rinse well. Incubate with the substrate working solution at room temperature for 2-10 minutes. Optimal development times should be determined by the investigator. Wash for 5 minutes in water. NOTES
- We recommend using glass-distilled water in the preparation of substrate buffer. Deionized water may con-tain inhibitors of the peroxidase reaction. Solutions containing sodium azide or other inhibitors of peroxidase activity should not be used in diluting the peroxidase substrate.
- DAB are carcinogens. Appropriate care should be exercised when using these reagents including gloves, eye protection, lab coats, and good laboratory procedures.
DAB Substrate Kit