Objective: to test IgG reactivity against SH-SY5Y neuroblastoma derived neurons in CNTN1/Caspr1+ patients
ICC protocol:
- Fix diferenciated SH-SY5Y cells with 4% PFA 5 min
- Wash 2 times with PBS 1x
- Block with goat serum 5% for 1h at RT
- Dilute serum 1/100 in blocking solution, incubate for 1h
- Wash with PBS 1x
- Commercial anti-NF Ab 1/200 diluted in blocking solution, incubate for 1h
- Wash with PBS 1x
- Secondary antibodies: GAH 488 (1/500), GAC 594 (1/500)
- Wash with PBS 1x (not water)
- Vectashield mounting medium
Results: see table and photos
| SAMPLE | RESULT |
|---|---|
| 18808 | NEGATIVE |
| 17595 | MILD POSITIVE |
| 18859 | POSITIVE ++ (SIMILAR TO CNTN1) |
| 19003 | MILD POSITIVE (PUNTEADO) |
| MRVM | MILD POSITIVE |
| 16047 | NEGATIVE (somas) |
| Vieira | MILD POSITIVE (axons and somas) |
| Piffrei | MILD POSITIVE |
| Regine | MILD POSITIVE |
| CNTN1+ | POSITIVE |
| Negative control | NEGATIVE |
