Objective: Confirmation of IgM positivity towards NF186 and NF140 of CMT and CIDP patients.
SAMPLES:
For NF186 and NF140 IgM:
| CODE | NF186 | NF140 |
|---|---|---|
| 18-532 (1) | - | - |
| 18-532 (2) | (few cells) | - |
| 18-533 | +/- (with doubts) | - |
| 9 | - | - |
| 17 | - | - |
| 34 | - | - |
| 38 | - | - |
| 43 | - | |
| 45 | - | |
| 55 | - | |
| 56 | - | |
| 57 | - | |
| CTRL - | - |
For NF186 and NF140 IgG:
| CODE | NF186 | NF140 |
|---|---|---|
| 23 | + | + |
| 57 | - | - |
| CTRL - | - | |
| CTRL+ | + |
NF140 and NF186 ICC:
- HEKs 293 Cells Transfection: 2018.08.14
- Blocking solution: 5% goat serum in PBS.
- Patient’s sera diluted 1/100 for IgG and 1/40 for IgM with 5% goat serum in PBS.
- Primary antibody: AF3235 diluted 1/1000.
- Secondary antibodies: For IgG: GAC 488 (1/1000) + GAH 594 IgG (1/1000) or GAH IgM (1/1000)
- Vectashield mounting medium.
RESULTS: All samples have an inespecific staining without colocalization; including the negative control; except the sample 23 for IgG towards NF140/NF186; tat seems to be positive (although with a lot of background). We are going to perform an ELISA with NF140 protein.
