DAB Substrate Kit – 60 ml – ab64238

DAB Substrate Kit – 60 ml – ab64238

• Product name

  DAB Substrate Kit

• Product overview

  DAB substrate kit ab64238 is used for IHC staining using HRP / peroxidase enzyme for detection. Reagent is stable for up to 6 hours after mixing the two components.

  DAB (3,3’Diaminobenzidine) is the most commonly used chromogen for immunohistochemical staining. In the presence of HRP / peroxidase, DAB produces a brown precipitate that is insoluble in alcohol.

  This product is a two component form consisting of a liquid, refrigerator stable DAB Chromogen and DAB Substrate.

  IHC protocol suitable for use with DAB substrate kit:
  For frozen sections, skip steps 1 and 2. 
  

  1. Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.

  2. Use appropriate antigen retrieval buffer or enzyme (primary antibody dependent) to treat sections. Wash 3 times in buffer.

  3. Add enough hydrogen peroxide blocking solution to cover the sections. Incubate for 10 minutes. Wash 2 times in buffer. If necessary, use avidin biotin blocking.

  4. Apply protein block (or normal serum from same species as secondary antibody) and incubate for 5 minutes at room temperature to block nonspecific background staining. Wash once in buffer.

  5. Apply primary antibody in antibody diluent and incubate.

  6. Wash 4 times in buffer. Incubate slide with biotinylated secondary antibody (or HRP polymer secondary antibody and skip step 7). Wash 4 times in buffer.

  7. Apply streptavidin-HRP and incubate for 10 minutes at room temperature.

  8. Rinse 4 times in buffer. Place slide in DAB substrate ab64238 or AEC substrate and incubate until desired color is achieved (1-10 mins). Rinse 4 times in buffer.

  9. Add enough drops of hematoxylin to cover the section. Incubate for 1 minute.

  10. Rinse 7-8 times in tap water. Add mounting medium to cover the section.

• Notes

  Find complete IHC kits, and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the IHC kits and reagents guide.

• Tested applications

  Suitable for: IHC-P