IHC Protocols for Mouse Anti-human RANTES
GUIDELINE
Immunohistochemistry (IHC) for mouse anti-human RANTES is a technique used to detect and visualize the presence of RANTES (regulated on activation, normal T cell expressed and secreted) in tissue samples. RANTES is a chemokine that plays a role in inflammation and immune response.
This specific immunohistochemistry method involves using a mouse anti-human RANTES antibody to bind to RANTES molecules in the tissue. The antibody is labeled with a marker, typically a dye or an enzyme, which provides a visual signal or color reaction when it binds to the RANTES.
METHODS
- Deparaffinize and rehydrate the tissue section.
- Perform heat-induced antigen retrieval with 0.1 M citrate buffer pH 6.0 at 123°C for 3 minutes in a pressure chamber. Wash the slide twice for three minutes each (50 mM Tris-HCl pH 8.0, 150 mM 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.
Creative Bioarray Relevant Recommendations
- Creative Bioarray offers a comprehensive IHC service from project design, and marker selection to image completion and data analysis. We are dedicated to satisfying every customer and assisting them to achieve their specific research goals.
- We provide various members of the RANTES (regulated on activation, expressed by normal T cells, presumably secreted) family, which are key members of the chemokine beta (CC) family.
NOTES
- The type of tissue being analyzed and the fixation method used can impact the IHC protocol. Different tissues may require specific antigen retrieval methods, and the choice between paraffin-embedding or frozen sections will influence the deparaffinization and rehydration steps.
- For formalin-fixed, paraffin-embedded tissues, antigen retrieval is crucial to unmask the target antigen and enhance antibody binding. The appropriate retrieval method (e.g., heat-induced epitope retrieval using citrate buffer) and conditions need to be determined.
- The selection of the mouse anti-human RANTES antibody is critical. It should be validated for IHC applications to ensure specificity and sensitivity for the target protein in the chosen tissue samples.
