Hello, I have a question about the Annexin V apoptosis assay. We have been doing this assay in our lab using lymphocytes and it has been working quite well. Now we are trying to do the same assay using an adherent breast cancer cell line (MDA-MB-231) and we are having some difficulties. Has anyone ever done the Annexin V assay using adherent cells? We are not sure if we should be doing the assay while the cells are still adhered to the plate or if we should harvest the cells prior to adding the Annexin V. Also, we cannot induce apoptosis in our adherent cells using our usual method (these tumor cells are Fas resistant) and we have not been able to find a method for doing this. Does anyone have any suggestions? Perhaps the Annexin V assay is not the best option for measuring apoptosis of adherent cells? If anyone could help us with these questions we would greatly appreciate it. Thank you, Sincerely, Lindsay Robinson Graduate Student Dept Agricultural, Food and Nutritional Science 4-10 Agriculture/Forestry Centre University of Alberta Edmonton, Alberta T6G 2P5 Phone: (780) 492-4240 Fax: (780) 492-9130
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