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Procedure for the early detection of apoptosis using annexin V staining and optional propidium iodide (PI).
The appearance of phosphatidylserine (PS) residues (normally hidden within the plasma membrane) on the surface of the cell is an early event in apoptosis and can be used to detect and measure apoptosis.
During apoptosis, PS is translocated from the cytoplasmic face of the plasma membrane to the cell surface. Annexin V has a strong, Ca2+-dependent affinity for PS and therefore can be used as a probe for detecting apoptosis.
Save time with a validated Annexin V assay kit/reagent
For robust results, we recommend our range of one-step Annexin V staining/detection kits and reagents for use in flow cytometry and cell imaging.
Alternatively, we would recommend the following protocol:
Analyze annexin V-FITC binding by flow cytometry (Ex = 488 nm; Em = 350 nm) using FITC signal detector (usually FL1) and PI staining by the phycoerythrin emission signal detector (usually FL2). For adherent cells, gently trypsinize and wash cells once with serum-containing media before incubation with annexin V-FITC (Steps A. 3–5).
The above is an example protocol for PS exposure detection using annexin V, based on the protocol provided in Annexin V-FITC Apoptosis Detection Kit (ab14085). Please be aware that, when using a kit, you should always use the protocol provided as it has been specifically designed for the product.