20170425: IgM primary culture Schwann cell ICC in Zika-GBS

Primary culture  Schwann cell IgM ICC is performed to test Zika-GBS patients’ staining pattern.

PATIENTS

The following patients  were analyzed by IgM:

Materials

• Rat primary culture of Schwann  grown in coverslips. Fixed with PFA 4% and blocked with GOAT 5% in PBS.
• PBS
• Blocking: GOAT 5% in PBS
• Patient’s sera diluted 1/40 with GOAT 5% in PBS
• Secondary Abs: GAH 488 IgM (1/1000) (box 21)
• Vectashield mounting medium

RAT PRIMARY CULTURE SCHWANN CELL ICC 

1. Primary culture Schwann cells  grown in coverslips were fixed and blocked with GOAT 5% in PBS and frozen at -80ºC.
2. Thaw these cells with a little GOAT 5% in PBS
3. Perform ICC:
   1. Add patient’s sera 1/40 1h at RT.
   2. Wash 3 times with PBS.
   3. Add secondary Ab GAH 488 IgM (1/1000) each and incubate 1h at RT.
4. Wash 3 times with PBS.
5. Mount with Vectashield mounting medium.

RESULTS