Objectives: To test samples for positivity towards FLOT1/FLOT2 (with live ICC).
Samples: Negative control, 158, 163.
Transfection: 03/09/2019 (48h)
FLOT1/FLOT2 ICC:
- Take 3 coverslips from the plate transfected the day before and put them in a 24-well plate
- Patient’s sera diluted 1/100 with HEK medium –> put 2,5 microl of serum in 250 microl of medium in every well. Incubate for 1h at 37ºC
- Remove medium and fix with PFA 4 % for 10 min.
- Blocking solution: Goat serum 5% diluted in PBS (1h).
- Primary antibody: FLOT1 1/100
- Secondary antibodies: GAR488 + GAH 594 1/1000 (1h)
- Vectashield mounting medium.
RESULTS: All samples seem positive, including the negative control
