Annexin V-iFluor 647 Apoptosis Staining / Detection Reagent

Annexin V-iFluor 647 Apoptosis Staining / Detection Reagent (ab219907) is a cell- impermeable reagent designed to bind to phosphatidylserine (PS) residues exposed on the outer cell surface of cells with a flow cytometer or fluorescence microscopy at Ex/Em = 651/660 nm.

We recommend using an impermeable nuclear stain such as Propidium Iodide (Propidium Iodide ab14083) or DRAQ7™ (DRAQ7™ ab109202) together with Annexin V-iFluor 647 Detection Reagent to discriminate necrotic and dead cells: plasma membrane is disrupted in these cells and therefore the Annexin V reagent will bind to PS found in the interior of cells.

Apoptosis is a regulated process of cell death that occurs during embryonic development as well as maintenance of tissue homeostasis. Inappropriately regulated apoptosis is implicated in different disease states, such as neurodegeneration disease and cancer. The apoptosis program is characterized by morphologic features, including loss of plasma membrane asymmetry and attachment, condensation off the cytoplasm and nucleus, and compaction and fragmentation of the nuclear chromatin. Exposure of phosphatidylserine (PS) on the external surface of the cell membrane has been reported to occur in the early phases of apoptotic cell death, during which the cell membrane remains intact. In leukocyte apoptosis, PS on the outer surface of the cell marks the cell for recognition and phagocytosis by macrophages. The human vascular anticoagulant, annexin V, is a 35-36 kDa Ca²⁺ dependent phospholipid binding protein that has a high affinity for PS, and shows minimal binding to phosphatidylcholine and sphingomyelin. Changes in PS asymmetry, which can be analyzed by measuring annexin V binding to the cell membrane, are generally observed before morphological changes associated with apoptosis occurred and before membrane integrity is lost.