Mouse T Cell Activation with anti-CD3ε Antibodies (clone 145-2C11)
Prepare a 5 µg/ml solution of anti-CD3ε (clone 145-2C11) 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 microwell plate.
Wash plate microwells 3 times with sterile PBS (wash 1/3).
Prepare single cell suspension of cells of interest.
Resuspend cells 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 (wash 2/3).
Wash plate microwells 3 times with sterile PBS (wash 3/3).
Discard liquid.
