Diaminobezidine (DAB) Kit

Mixing of reagent: 1. To five ml of distilled or deionized water in a test tube, add two drop of reagent B, mix Well. 2. Add one DAB tablet (Reagent C), let stand in this solution for 5-10 minutes. Vortex for 3-5 minutes till all DAB tablet is dissolved, add one drop of reagent S, and mix Well. 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, incubate at RT or 37ºC for 5-10 minutes. 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-6520) or any counterstain. 5. Wash with tap water, buffer pH 7.4 or over, followed by rinsing with distilled/deionized water. 6. The slide can be mounted with aqueous mounting medium or organic mounting medium. For organic mounting medium dehydration of slide is required.

Reagent B: concentrated Buffer 5 ml (yellow color cap) Reagent C: DAB chromogen tablets 20 (amber color cap) Reagent S: substrate, H2O2 3 ml (white color cap)

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.