DAB (3, 3 -diaminobenzidine) HRP substrate produces a dark brown reaction product and can be used for both immuno-histochemical and blotting applications. This substrate kit is an optimized formula that is crisper and is 3 - 4 times more sensitive than conventional DAB substrate kit.
PREPARATION OF SUBSTRATE WORKING SOLUTION:
Add 1 drop (approximately 30 µL) of DAB Chromogen concentrate to 1 mL DAB Diluent. Mix well before use.
INSTRUCTIONS FOR USE
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 or PBS. Counterstain if desired.
NOTE: Unused working solution is stable up to 14 days if stored at 2°- 8ºC or up to 5 days if stored at room temperature (approximately 23ºC). Working solution may change color during storage but this will have no effect on the quality or intensity of the staining. No sample-obscuring precipitate will form. It is not necessary or recommended to add detergent to ImmPACT™ DAB working solution to reduce surface tension (e.g. for use in an automated stainer). The DAB reaction product can be intensified using a DAB Enhancing Solution after development.
IMPORTANT SAFETY INFORMATION
DAB is a suspected carcinogen. Appropriate care should be exercised when using this reagent including gloves, eye protection, lab coats, and good laboratory procedures. Dispose in accordance with local regulation.
Store reagents in original bottles at 2°- 8ºC. Avoid storing reagents or working solution in strong direct light.