Samples:
- N31 (control)
- N33 (control)
- N34 (control)
- N35 (case)
- N36 (case)
- N37 (case)
Immunohistochemistry (in brain frozen sections):
- Fix with acetone (5 min)
- Wash with PBS 1x (3 x 5 min)
- Block with Goat Serum 5% + 2 % BSA (10 ml Goat serum 5% in PBS + 0’2g BSA)
- Primary antibody: rabbit anti-NF155 (15035, Cell Signal). Diluted 1/50 in blocking goat (1h)
- Wash with PBS 1x (3 x 5 min)
- Secondary antibodies (1 h): GAH488 IgG+ GAR594 diluted 1/1000 in blocking goat (1h)
- Wash with PBS 1x (3 x 5 min)
- Fluoromount mounting medium
RESULTS:
- The pan-neurofascin doesn’t stain any specific structure.
- In the 3 cases there is a green staining in Purkinje cells (in comparison with the 3 controls) –> the human Ig is detected.
