Lindsay, Try doing the assay in non-stick polypropyrene tubes (eg Falcon Cat. No. 2063). John ------------------------------------------------------------------------ Dr. John N. Waitumbi c/o Dr Jose A. Stoute, USAMRU, Unit 64109, Box 401 APO AE 09831-4109 Kenya Phone +254 35 22942 Email:waitumbi@net2000ke.com Fax +254 35 22903 ---------- > From: Lindsay Robinson <robinson@gpu.srv.ualberta.ca> > To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> > Subject: Annexin V > Date: Friday, June 25, 1999 2:40 PM > > > 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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