Dear Everyone: 1. I transiently cotransfected into a human multiple myeloma cell line (that is very difficult to transfect) a GFP neo expression vector along with an expression vector in 1:10 and 1:20 ratio and then determine the effect of overexpression of specific protein on drug-induced apoptosis. Since the transfection efficiency is low (2-10%), I wanted to gate on the GFP population and then measure annexin V binding using PE-annexin.. I would like input re: optimal parameters used to aquire as well as analyze data. Is my strategy correct? 2. Does anyone have experience using mitocapture mitochondrial apoptosis detection kit (Alexis Biochemicals)? If so, how do perform the analysis. Thank you in advance, Rena Rena Feinman, PhD Assistant Professor of Surgery, UMDNJ-New Jersey Medical School
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