Flow cytometric analysis of Jurkat cells, untreated (left) or treated with camptothecin (10 uM, 4 hr; right), using Annexin V-PE Early Apoptosis Detection Kit.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
posted May 2019
Protocol Id: 1824
The Annexin V-PE Early Apoptosis Detection Kit enables researchers to identify early apoptotic cells within a cell population. Annexin V-PE conjugated protein binds to cell surfaces expressing phosphatidylserine, an early apotosis marker. Cells stained with 7-Aminoactinomycin D (7-AAD), a non-cell-permeable DNA dye, indicate necrotic cells. Cells stained with both 7-AAD and annexin V-PE demonstrate later stage apoptosis and early necrosis. This kit provides enough reagent to perform 100 assays, based on a 250 μl assay volume.
Annexin V belongs to the annexin family of proteins consisting of over 160 members. All annexin family members share the same characteristic of Ca2+-dependent binding to negatively charged phospholipid surfaces (1). During early apoptosis, phosphatidylserine (PS) is translocated from the cytosolic side of the plasma membrane to the cellular surface. This translocation exposes PS to the extracellular environment with the plasma membrane left intact (2). Annexin V protein demonstrates high affinity, specificity, and sensitivity for PS and can be used as a marker of early apoptosis. In order to rule out "leaky" necrotic cells, annexin V must be used in tandem with reagents that determine the integrity of the cell membrane, such as propidium iodide (2).
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