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 appropriateantigen 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, useavidin biotin blocking.

4. Applyprotein block(or normal serumfrom 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 inantibody diluentand incubate.

6. Wash 4 times in buffer. Incubate slide withbiotinylated secondary antibody(orHRP polymer secondary antibodyand skip step 7). Wash 4 times in buffer.

7.Applystreptavidin-HRPand incubate for 10 minutes at room temperature.

8. Rinse 4 times in buffer.Place slide in DAB substrate orAEC substrateand incubate until desired color is achieved (1-10 mins). Rinse 4 times in buffer.

9. Add enough drops ofhematoxylinto cover the section. Incubate for 1 minute.

10. Rinse 7-8 times in tap water. Addmounting mediumto cover the section.

Find completeIHC kits, and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in theIHC kits and reagents guide.