20181119: IgG and IgM primary culture Schwann cell ICC in controls

Primary culture Schwann cell IgG and IgM ICC is performed to test controls staining pattern.

Samples: 2 sera from healthy controls, 5 sera from ALS patients, and 5 sera from dysferlinopathy patients.

Protocol:

1. Primary culture Schwann cells  grown in coverslips were fixed with PFA 4 % 10 min,  blocked with Goat 5% in PBS 1h, and frozen at -80ºC.
2. Thaw these cells with a little Goat 5% in PBS
3. Patient’s sera 1/40 for IgM and 1/100 for IgG 1h at RT
4. Wash 3 times with PBS.
5. Secondary Ab GAH 488 IgG or IgM (1/1000) 1h at RT
6. Wash 3 times with PBS.
7. Mount with Vectashield mounting medium.

Results: