Department of Neurobiology and Behavior

Gunma Universtiy School of Medicine

1. Incubate the sections in PBS 15 min for removing OCT compound

(2. option: Incubate the sections in 0.1% Triton X-100 for 10 min and next in PBS for 10 min.)

(2' For N-CAM staining, incubate the sections in absolute methanol for 30 min.)

3. Incubate the sections in PBSA3 (3% BSA in PBS) for 30 min

4. Incubate the sections with first antibody for 2 hours. For control sections, skip this step.

5. Wash the sections 10 min three times with PBS.

6. Incubate the sections with FITC-conjugated goat anti-mouse IgG (H & L) 1:100 dilution in PBSA for 30 min.

7. Wash the sections in PBS for 10 min three times

8. Mount the sections with Perma flow.