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1. Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.
2. Add enough drops of an appropriate Hydrogen Peroxide Block to cover the sections. Incubate for 10 minutes. Wash 2 times in buffer.
3. Perform appropriate pretreatment if required. Wash 3 times in buffer.
4. Apply an appropriate Protein Block and incubate for 5 minutes at room temperature to block nonspecific background staining. Wash 1 time in buffer.
5. Apply primary antibody and incubate according to manufacturer's protocol.
6. Wash 4 times in buffer. Incubate slide with biotinylated secondary solution according to manufacturer’s protocol. Wash 4 times in buffer.
7. Apply Streptavidin Peroxidase and incubate for 10 minutes at room temperature.
8. Rinse 4 times in buffer. Place slide in appropriate chromogenic substrate and incubate until desired reaction is achieved. 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.
Our Abpromise guarantee covers the use of ab64269 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent dilution.|
|Sandwich ELISA||Use at an assay dependent dilution. PubMed: 21371998|