Sample: N33 (control)
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) (1 h)
- Primary antibody: anti-calbindin (ab82812 Abcam) diluted 1/1000 in blocking goat (1h)
- Wash with PBS 1x (3 x 5 min)
- Secondary antibody: GAM 488 diluted1/200 in blocking goat (1 h)
- Wash with PBS 1x (3 x 5 min)
- Fluoromount mounting medium
Result: The staining in Purkinje cells is very low. There is an inespecific staining in all the vessels of the brain.
Maybe this inespecific staining is due to the crossreaction between the secondary antibody anti-mouse and the rat brain.
