Objective: to test IgG reactivity against SH-SY5Y neuroblastoma derived neurons in patients iwht small fiber neuropathy
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 1/40 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 IgM (1/500), GAC 594 (1/500)
- Wash with PBS 1x (not water)
- Vectashield mounting medium
Results: see table and photos
| IVIG SAMPLE | RESULT |
|---|---|
| 1 | negative |
| 2 | POSITIVE ++ |
| 3 | positive: axons |
| 4 | negative |
| 5 | negative |
| 6 | positive: axons |
| 7 | negative |
| 8 | negative |
| 9 | positive: axons |
| 10 | positive: axons |
| 11 | negative |
| 12 | negative |
| 13 | positive: axons |
| 14 | positive: axons |
| 15 | negative |
| 16 | negative |
| 17 | positive: axons |
| 18 | positive: axons |
| 19 | negative |
| 20 | negative |
| 21 | negative |
| 22 | negative |
| 23 | negative |
| 24 | positive: axons |
| 25 | negative |
| 26 | negative |
| 27 | negative |
| 28 | negative |
| 29 | positive: somas + axons |
| 30 | positive: axons |
| 31 | negative |
| 32 | negative |
| 33 | negative |
| 34 | negative |
| 35 | negative |
| 36 | negative |
| 37 | negative |
| 38 | negative |
| 39 | negative |
| 40 | negative |
| 41 | negative |
| 42 | negative |
| 43 | negative |
| 44 | positive: axons |
| 45 | negative |
| 46 | positive: axons (mild) |
| 47 | positive: axons |
| 48 | negative |
| 49 | negative |
| 50 | positive: axons |
| 51 | positive: axons |
| 52 | |
| 53 | |
| 54 | |
| 55 | |
| 54 | |
| 56 | |
| 57 | |
| 58 | |
| 59 | |
| 60 |
Only sample 2 was clearly postive. The rest showed mild positivy against some axons. This sample was also IgG positive, but IgM positivity was stronger
