Human T Cell Activation with anti-CD3 Anitbodies (clone UCHT1, OKT3 or HIT3a)
Prepare a 10 µg/ml solution of anti-CD3 (clone UCHT1, OKT3, or HIT3a) in sterile PBS.
Dispense 50 µl of the antibody solution to each microwell of the 96-well assay plate.
For the unstimulated control wells, add 50 µl of sterile PBS.
Seal plate.
Incubate at 37°C for 2 hours or 4°C overnight.
Aseptically decant antibody solution from the microwell plate.
Wash plate microwells 3 times with sterile PBS (was 1/3).
Prepare single cell suspension of cells of interest in supplemented cell culture medium to 1-2 x 106/ml.
Aliquot 200 µl cell suspension into plate microwells.
Cover with lid.
Incubate at 37°C in 5% CO2 and 100% humidity for 3 days.
Wash plate microwells 3 times with sterile PBS (was 2/3).
Wash plate microwells 3 times with sterile PBS (was 3/3).
Discard liquid.
