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Protocols - Immunofluorescence: Rabbit Anti-Murine VEGF

The following protocol used normal murine kidney tissue with an autostainer.
  1. Deparaffinize and rehydrate the tissue section.
  2. Perform heat-induced antigen retrieval with a high pH buffer for 20 minutes.  Allow the slide to cool for 20 minutes.
  3. Wash the slide for 5 minutes (TBS/Tween 20).
  4. Use the following autostainer protocol:
         a. Incubate the tissue section for 1 hour at room temperature with Rabbit Anti-Murine VEGF at 2.5 μg/mL.
         b. Wash the slide.
         c. Incubate the tissue section with a fluorescent conjugated secondary antibody for 30 minutes at room temperature.
         d. Wash the slide.
  5. Counterstain the tissue section with DAPI.