First Edition (Revised on April, 2016)
DAB (3,3N-Diaminobenzidine Tertrahydrochloride), is one of the most important chromogens for peroxidase. It will form a dark brown precipitate in the presence of peroxidase enzyme. DAB Chromogen Kit is a widely used chromogen for immunohistochemical staining and immunoblotting. This product consists of a liquid stable DAB chromogen and DAB substrate buffer.
REAGENTS AND MATERIALS PROVIDED
|Chromogen A (1×)||10mL||Buffer|
|Chromogen B (50×)||200μL||DAB|
STORAGE AND PERIOD OF VALIDITY
Chromogen A should be stored at -20°. Chromogen B should be at 4℃for short time storage, while long time storage at -20°. Protect the reagents or working solution from the light. Do not use after expiration date printed on the kit.
1. After incubation of labelled antibody with peroxidase, wash the tissue sections with TBS or PBS for 4 times.
2. Mix 1mL Chromogen A with 20μL Chromogen B thoroughly, and then add this solution to the sample. And incubate the section for 30sec to 10mins.
3. Use the distilled water to end the reaction. Counterstain the sample with haematoxylin for 1~2 minutes, 1% HCl in ethanol for 30 seconds, then wash with distilled water, and followed by incubation in ammonia for 10 minutes.
4. Dry, brighten, embed with neutral resins, and then observe under a microscope.
DAB is a suspected carcinogen. Appropriate care should be exercised when using this reagent, including gloves, glasses, lab coats, and good laboratory procedures.