Hydrogen Peroxide Blocking Reagent

Hydrogen Peroxide Blocking Reagent ab64218 for use in IHC with detection based on HRP / peroxidase.

Note: The concentration of hydrogen peroxide in this product is 0.3%.

IHC protocol suitable for use with Hydrogen Peroxide Blocking Reagent:
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 ab64218 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 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.

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