T Cell Activation with anti-CD3 Antibodies Protocol - Mouse
- Prepare a 5µg/ml solution of anti-CD3ε (clone 145-2C11) in sterile PBS.
- Dispense 50µl of the antibody solution to each microwell of the 96-well assay plate. For the unstimulated control wells, add 50µl 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. Discard liquid.
- Prepare single cell suspension of cells of interest.
- Resuspend cells in supplemented cell culture medium to 1-2 x 106/ml.
- Aliquot 200µl cell suspension into plate microwells. Cover with lid. Incubate at 37°C in 5% CO2 and 100% humidity for 3 days.
General Tips & FAQ
- What if I want to stimulate my cells to a particular T-cell lineage?
- As a guide for this, you can take a look at our Activation Bundles page, which will provide you with the listing of our products you will need to differentiate into different T-helper cell lineages.