Annexin V-iFluor 488 Apoptosis Detection Kit

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Annexin V-iFluor 488 Apoptosis Detection Kit (ab219916) contains Annexin V labeled with our proprietary green fluorescent dye iFluor 488, which allows the identification and quantitation of apoptotic cells on a single-cell basis by flow cytometry. Optionally, it can be used for simultaneous staining of cells with Annexin V-iFluor488 (green fluorescence) and the nonvital dye propidium iodide (PI) (orange fluorescence) which allows the discrimination of intact cells (Annexin V-iFluor 488 negative, PI Staining Solution negative), early apoptotic (Annexin V-iFluor 488 positive, PI Staining Solution negative) and late apoptotic or necrotic cells (Annexin V-iFluor 488 positive, PI Staining Solution positive).

The iFluor 488 dye (Ex/Em = 490/525 nm) has spectral properties almost identical to those of FITC or Alexa Fluor® 488, making it convenient to be used with the common fluorescence instruments equipped with the light sources and filters for FITC, the most common fluorophore.

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.

Alexa Fluor^® 488 is the trademark of Invitrogen.