Objective: To confirm positivity for NF140/186 in 2 samples, positive for ICC but negative in ELISA. We are going to perform it in non-transfected cells; with and without the commercial; to detect cross-reactivity.
Samples: 18-720, 18-806, neg ctrl and pos ctrl.
ICC NF155/NF140/NF186:
- Blocking solution: 5% goat serum in PBS.
- Patient’s sera diluted 1/100 in 5% goat serum in PBS.
- Primary antibody: AF3235 diluted 1/1000.
- Secondary antibodies: GAC488 + GAH 594 IgM (1/1000).
- Vectashield mounting medium.
RESULTS:
The samples were negative in non-transfected cells.
Sample 18-720 is negative with and without commercial.
Sample 18-806 is positive with commercial and clearly positive for NF155 and NF186 without commercial antibody. The NF140 is not clearly positive, but the positive control is negative too. Sample 18-803 is negative for NF140 ELISA and it is negative for teasing too. We are going to perform ELISA NF155.
