Protocol:
- Cells seeded on 06.02.20, differentiated on 07.02.20 and 10.02.20
- Fix for 15 minutes with PFA 4 %
- Wash 3 times with PBS1x
- Block for 1 hour with Goat Serum 5 %
- Patient’s sera
- For IgG: diluted 1/100 in Goat serum 5 % (1 h)
- Wash 3 times with PBS1x
- Commercial antibody: anti-NF (AF3235) diluted 1/200 in Goat serum 5% (1h)
- Wash 3 times with PBS1x
- Secondary antibodies: GAC594 IgG + GAH488 IgG diluted 1/500 in Goat serum 5 % (1 h)
- Wash 3 – 6 times with PBS1x (NO H2Od)
- Vectashield mounting medium
Samples and results:
