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 .
