Description
Product details:
AEC Substrate kit (ab64252) for use in IHC using HRP / peroxidase detection. Ready to use formulation.
Previously called AEC Substrate System (Ready to Use).
IHC protocol suitable for use with AEC Substrate Kit (ab64252)
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
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
AEC Substrate ab64252 and incubate until desired color is achieved (1-10 mins, recommended 10 mins for AEC)
. 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
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-Please refer to protocols
