The following protocol used normal murine kidney tissue with an autostainer.
Deparaffinize and rehydrate the tissue section.
Perform heat-induced antigen retrieval with a high pH buffer for 20 minutes. Allow the slide to cool for 20 minutes.
Wash the slide for 5 minutes (TBS/Tween 20).
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.