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Protocols - Immunofluorescence: Mouse Anti-Human IL-6

The following protocol used human endometrial carcinoma tissue.

  1. Deparaffinize and rehydrate the tissue section.  Wash the slide in DI and then in wash buffer (1X PBS/ 0.05% Tween 20).
  2. Perform heat-induced antigen retrieval with a steamer and incubate the tissue section in 10mM pH 6.0 sodium citrate buffer for 20 minutes.  Allow the slide to cool for 20 minutes.  Wash the slide twice for three minutes each.
  3. Incubate the tissue section with an Fc receptor block for 20 minutes.  Wash the slide twice for three minutes each.    
  4. Incubate the tissue section with a protein blocking buffer for 10 minutes.   
  5. Incubate the tissue section overnight at 4˚C with Mouse Anti-Human IL-6 at 0.125 μg/mL.  Wash the slide twice for three minutes each.
  6. Incubate the tissue section with a fluorescent conjugated secondary antibody for  2 hours at room temperature. Wash the slide twice for three minutes each.
  7. Counterstain the tissue section with DAPI.