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Annexin V Binding Buffer is recommended for use with Annexin V reagents. Cells should be resuspended in Annexin V Binding Buffer at a concentration 1x106/ml. The binding buffer contains an optimal concentration of calcium which is required for Annexin V binding to phosphotidylserine (PS) on the cell surface. Annexin V is a calcium dependent phospholipid binding protein that preferentially binds to negatively charged phospholipids including PS. Cells progressing through apoptosis can be monitored according to their Annexin V and propidium iodide staining pattern. Early apoptotic cells will bind Annexin V but are not sensitive to intracellular staining with propidium iodide (PI). As cells progress through apoptosis the integrity of the plasma membrane is lost, allowing PI to penetrate and label the cells with a strong yellow-red fluorescence.
1. Vermes, I. 1995. J. Immunol. Methods. 184(1): 39-51. PMID: 7622868
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