- Harvest 2x105 cells per staining and wash cells twice in cold phosphate- buffered-saline (PBS).- Resuspend cells in 100µl binding buffer (10mM HEPES, pH 7.4, 140mM sodium chloride, 2.5mM calcium dichloride), add 5µl annexin V-Alexa Fluor® 488 to the cells and mix gently. For detection of necrosis add additionally appropriate amounts of propidium iodide (final concentration 1µg/ml or 1.5µM).- Incubate for 15 min. at room temperature in the dark.- Add 400µl binding buffer and analyze the staining in flow cytometry as soon as possible (within an hour).
Swiss-Prot link P48036: Annexin V (mouse)AfCS Signalling Gateway link A000281: Annexin V (mouse)