Objectives: To test CMT patients for IgG/IgM positivity towards NF155 and CNTN1.
Samples:
For NF155 IgM:
| CODE | RESULTS |
|---|---|
| 9 | - |
| 10 | - |
| 11 | - |
| 12 | - |
| 13 | - |
| 48 | - |
| 49 | - |
| 50 | - |
| 51 | - |
| 52 | - |
| 53 | NO REALIZADO |
| 54 | - |
| 55 | - |
| 56 | - |
| 57 | - |
For NF155 IgG:
| CODE | RESULTS |
|---|---|
| 23 | + |
| 34 | - |
| 49 | - |
| 50 | - |
| 51 | - |
| 52 | - |
| 53 | - |
| 54 | - |
| 55 | - |
| 56 | - |
| 57 | - |
| CTRL + | + |
| CTRL - | - |
For CNTN1 IgM:
| CODE | RESULTS |
|---|---|
| 12 | - |
| 24 | - |
| 49 | - |
| 50 | - |
| 51 | - |
| 52 | - |
| 53 | - |
| 54 | - |
| 55 | - |
| 56 | - |
| 57 | - |
| CTRL + | + |
| CTRL - | - |
For CNTN1 IgG:
| CODE | RESULTS |
|---|---|
| 9 | - |
| 10 | - |
| 11 | - |
| 12 | - |
| 13 | - |
| 14 | - |
| 15 | - |
| 16 | - |
| 17 | - |
| 18 | UNCERTAIN (REPEAT) |
| 19 | - |
| 20 | - |
| 21 | - |
| 22 | - |
| 23 | - |
| 24 | - |
| 25 | - |
| 26 | - |
| 27 | - |
| 28 | - |
| 29 | - |
| 30 | - |
| 31 | - |
| 32 | - |
| 33 | - |
| 34 | - |
| 48 | - |
| 49 | - |
| 50 | - |
| 51 | - |
| 52 | - |
| 53 | - |
| 54 | - |
| 55 | - |
| 56 | - |
| 57 | - |
| CTRL - | - |
NF155 ICC:
- HEKs 293 Cells Transfection: 2018.01.16
- Blocking solution: 5% goat serum in PBS.
- Patient’s sera diluted 1/40 for IgM and 1/100 for IgG 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)
- For IgM: GAC 488 (1/1000) + GAH 594 IgM (1/1000)
- Vectashield mounting medium.
CNTN1 ICC:
- HEKs 293 Cells Transfection: 30.05.2018
- Blocking solution: Rabbit serum diluted 1/40 in PBS.
- Patient’s sera diluted 1/100 for IgG with rabbit serum 1/40.
- Primary antibody: AF904 diluted 1/1000.
- Secondary antibodies: RAG 488 + RAH 594 IgM (1/1000)
- Vectashield mounting medium.
Results: The sample 23 is positive for IgG towards NF155. However, it was positive for IgG towards NF140 too and we suspect than the staining can be inespecific. We can make a ICC with no transfected NEK cells to see if the staining is inespecific. The rest of the samples were negative for IgM or IgG towards NF155.
We have a problem with the primary antibody AF904 and we can’t co-localize a sample that was uncertain: sample 18 for IgM; so we are going to repeat it. There is an inespecific staining in sample 24 for IgG. The rest of the samples were negative.
