GFP Staining - KravitzLab/KreedLabWiki GitHub Wiki

First day

  1. Rince the brain slices in PBS 1X 3x10 minutes (Skip this step if the brain slices were store in PBS and not cryprotectant)
  2. Incubate the slices at room temperature with 5% of Donkey Serum in PBS-T 0.5% for about an hour.
  3. Incubate the slices at 4°c on an agitator with 1:1000 of chicken anti GFP antibody (Aves Lab antibody in the -20 fridge)

_Note : The antibody works very VERY well at 1:1000, it should probably work at 1:2000 dilution) _

Second day

  1. Rince the brain slices in PBS 1X 3x10 minutes.
  2. Incubate the slices at room temperature with 1:500 Goat anti chicken secondary antibody for 3 hours (Covered in aluminum foil)
  3. Rince the brain slices in PBS 1X 3x10 minutes
  4. Mount the brain slices

Example :

image