Diaminobezidine (DAB) Kit REMOVE

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.

Peroxidase chromogen-substrate 3-3´ diaminobezidine (DAB) reagent is an excellent chromogen system that is used in Immunohistochemistry (IHC) and Immunocytochemistry (ICC). This chromogen system gives chocolate brown reddish color and is compatible with hematoxylin (AR-6520), nuclear fast red (AR-6524), methylene blue (AR-6525) or Methyl green (AR-6526) counterstain. This chromogen with above counterstains are insoluble in alcohol and organic solvents (dehydrating agents); therefore can be mounted with organic mounting mediums; e.g. our Organo mounting medium (AR-6504) or others. DAB can also be mounted with aqueous mounting medium (AR-6503) if hematoxylin is used as counter stain.