Protocols - Immunohistochemistry: Mouse Anti-Human RANTES
The following protocol used formalin-fixed paraffin-embedded human tonsil tissue.
*Protocol and staining provided by Dr. Lauren Binge, Laboratory of Prof. Charles Mackay, Monash University, Australia
- Deparaffinize and rehydrate the tissue section.
- Perform heat-induced antigen retrieval with 0.1M citrate buffer pH 6.0 at 123°C for 3 minutes in a pressure chamber. Wash the slide twice for three minutes each (50mM Tris-HCl pH 8.0, 150mM NaCl in distilled water).
- Incubate the tissue section with 1% BSA/PBS block.
- Incubate the tissue section overnight at 4°C with Mouse Anti-Human RANTES at 10.0 µg/mL. Wash the slide twice for three minutes each.
- Incubate the tissue section with an HRP polymer kit according to the manufacturer’s protocol. Wash the slide twice for three minutes each.
- Incubate the tissue section with the DAB chromogen.
- Counterstain the tissue section with hematoxylin.
- Coverslip the sections with mounting medium.