20140530 ICC CNTN

Objective

The aim of this experiment is to test sera from 5 new patients and a suspicious negative control.

Materials

• Transfected HEK cells with CNTN1-human grown in coverslips
• Patients sera
• Antibodies:
  • mouse-anti-contactin
  • GAM-488
  • GAH-594

Procedure

1. Take one coverslip per condition (7)
2. Prepare sera 1:40 in goat serum. Add a drop (40µl) of each preparation to the parafilm. Incubate for 1h
3. Wash 3 times with PBS
4. Incubate primary antibody 1h: mouse-anti-contactin 1:500
5. Wash 3 times with PBS
6. Prepare secondary antibody: (A) GAH IgG-594 1:500 + GAM-488 1:500. Add a drop and incubate 1h.
7. Wash 3 times with H2O
8. Mount with Vectashield (with DAPI)

Results & conclusions

All patients are clearly negative including the suspicious negative control.