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.
|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. |
- 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.