Samples: N33 (control) and N36 (case)
Immunohistochemistry (in brain frozen sections):
- Fix with acetone (5 min)
- Wash with PBS 1x (3 x 5 min)
- Block with Blocking Goat (10 ml PBS + 0’5 ml Goat Serum + 0’2 g BSA) (30 min)
- Primary antibody: anti-calbindin. Diluted 1/500 in blocking goat
- Wash with PBS 1x (3 x 5 min)
- Secondary antibodies (1 h): GAH488 + GAM594 diluted 1/200 in blocking goat
- Wash with PBS 1x (3 x 5 min)
- Fluoromount mounting medium
RESULT:
- Ac anti-calbindin: marca bien las células de Purkinje (en el control y en el caso)
- GAH488: en el caso se observa marcaje en las células de Purkinje y en los tractos de mielina.
