Diaminobezidine (DAB) Kit

Mixing of reagent: 1. To one ml of Buffer Substrate (BS) in a test tube, add two drop of Chromogen (C) reagent mix well. 2. This ready-to-use reagent is good for several hours. The unused DAB solution can be discarded according to city, county, state, province or country’s regulations. Application: IHC, ICC 1. After the peroxidase reaction, wash slide with buffer (this buffer should not contain any sodium azide because peroxidase is inactivated) and distilled/ deionized water. 2. Add few drops of ready to use DAB reagent on tissue slides for 5-10 minutes at RT. (Higher temperature 37°C can also be used for DAB chromogen, however the incubation time should be determined by the individual lab) 3. Wash 2-5X with buffer, followed by rinsing with distilled/deionized water. 4. Add counterstain compatible with DAB, we use hematoxylin 30-60 seconds (cat. # AR-6521) or any counterstain. 5. Wash with tap water, buffer pH 7.4 or over, followed by rinsing with distilled/deionized water. 6. Mount slide with aqueous mounting medium or organic mounting medium, after dehydration as described in the protocol for mounting medium.