Did anybody use MoAb anti TCR alpha beta and gamma delta to block the receptor before performing cytotoxicity of effector lymphocytes against cancer cells? I'm going to use BioLegend Mo Ab against TCR but I haven't found any suggestions about the MoAb concentration to use and the time of incubation. Could someone help me? Thanks in advance.
Hi Paola,
In our lab, we usually block our chimeric antigen receptor (CAR) T cell cytotoxicity by pre-incubation CAR T cells with 2 to 5ug antibody/well for 30mins at 37C. Then you plate target cells into the plate. [Our general condition is 10^5 T cells+ + 2ug antibody +2x10^4 target cells in 200uL total volume in a well of 96 well plate.](You can also do it the reverse way, plate target cells, then using antibody against the ligand to pre-block ligand on tumor cells, then add T cells). Hope this helps.
thanks for your suggestions, I'll try to adapt your experimental scheme to my protocol.
- Badges
- Science method