1. 二抗带生物素/HRP：需要用DAP显色试剂盒才能显示颜色，例如abcam的DAB底物试剂盒(ab64238)
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.
Apply [streptavidin-HRP]and incubate for 10 minutes at room temperature.
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.
Add enough drops of [hematoxylin]to cover the section. Incubate for 1 minute.
Rinse 7-8 times in tap water. Add [mounting medium]to cover the section.
