Hi Kenishe, we normally assay annexin V binding on adherent cells. We harvest them with trypsin-EDTA and afterwards (I think that's the main point) we wash them twice in annexinV binding buffer before adding the annexin. It works quite well for almost all lines assayed, and control cells do not show high "apoptosis". ( E.Castaño, M.Dalmau, M.Barragán, G.Pueyo, R.Bartrons, J.Gil (1999) Aspirin induces cell-death and caspase-dependent phosphatidylserine externalization in HT-29 human colon adenocarcinoma cells. British J. Cancer 89,2: 294-299) Hope this helps. Mireia ________________________________________________________ Mireia Dalmau, PhD Flow Cytometry laboratory Campus de Bellvitge, Universitat de Barcelona. C/ Feixa Llarga s/n. 08907 L'Hospitalet Telf. 93 402 42 06 FAX. 93 402 42 13 e-mail: serv-rad-bell@bell.ub.es __________________________________________________________
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