Can anyone point out the reasons for inaccurate activation of nave T cells post negative MACS selection ? I had pre-coated wells of 48 well plate with 2ug/mL of antiCD3 abs and had it incubated for 2 hrs prior to seeding those cells. Thereafter, with the polarizing cytokines and soluble anti-CD28 ab (1 ug/mL) were added. I had seeded about 300,000 cells per well. It has been just one day post seeding, the cells still look smaller. What do I do ???
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