20180119: Check crossreactivity in NF140 IgM ICC

Objectives: To discard a crossreactivity between the comercial AF3235 and the secondary antibodies, suspected in the experiment of 2018/01/17.

Patient: 18-532

NF140 ICC:

• HEKs 293 Cells Transfection: 2017.11.22
• Blocking solution: 5% goat serum in PBS.
• Patient’s sera diluted 1/10 for IgM with 5% goat serum in PBS.
• Primary antibody: AF3235 diluted 1/1000.
• Secondary antibodies: GAH 594 IgM (1/1000) + GAC 488 IgY (1/1000).
• Vectashield mounting medium

We performed a ICC with different samples:

1. Sera 18-532 + GAH 594 IgM
2. AF3235 + GAC 488
3. Sera 18-532 + Secondary antibodies (GAH 594 IgM and GAC 488)
4. AF3235 + Secondary antibodies (GAH 594 IgM and GAC 488)

Results: We found a positive result in samples 1 and 3 and a negative result in samples 2 and 4; so we discard a crossreactivity between the comercial AF3235 and the secondary antibodies.