|FineTest 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 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℃|
- 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 2 drops (approximately 50 µl) of Reagents C
- Add 2 drops (approximately 50 µl) of Reagents A
- Add 2 drops (approximately 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.|
- 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.