Hi Dave, This is a head scratcher. Did the whole population move in response to the anti-CD3, or only a small fraction of the cells? I'd try using the anti-CD3 and a secondary to label the splenocyte culture, and find out how many CD3 positive cells you have. This should answer two questions- are the expected number of T-cells actually in this culture, and does this lot of antibody actually bind?. Peter Lopez The Aaron Diamond AIDS Research Center
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