Protein blocking buffer ab64226 for serum-free blocking of non-specific antibody binding in IHC, ELISA and western blot. Ready to use. No mixing or dilution required.

For use in ELISA, add Protein Block to well and incubate for 2-10 minutes before addition of sample. Wash and continue procedure. For use in western blotting, incubateforone hour at room temperature or overnight at 4°C.

IHC protocol suitable for use with Protein Block ab64226:For frozen sections, skip steps 1 and 2. For fluorescent IHC, skip step 3, incubate with fluorescent dye conjugated secondary at step 6, skip rest of steps, and mount with anti-fade mounting medium.

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 enoughhydrogen peroxide blocking solutionto cover the sections. Incubate for 10 minutes. Wash 2 times in buffer.If necessary, useavidin biotin blocking.

4. Apply Protein Block ab64226(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.Wash 4 times in buffer.

6. Incubate withbiotinylated secondary antibody(orHRP polymer secondary antibodyand 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 inDAB substrateorAEC substrateand incubate until desired color is achieved. 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.