Flow cytometric analysis of Jurkat cells untreated (left) or treated with camptothecin (10uM, 4 hr; right) using Annexin V-FITC Early Apoptosis Detection Kit.
Confocal immunofluorescent analysis of live Jurkat cells, untreated (left) or camptothecin-treated (right), using Annexin V-FITC Conjugate (green). Red = Propidium Iodide (fluorescent DNA dye). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
|Product Includes||Quantity (with Count)|
|Annexin V-FITC Conjugate||1 x 100 µl|
|10X Annexin V Binding Buffer||2 x 1.5 ml|
|Propidium Iodide (PI) Solution||1 x 1.3 ml|
Note: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
posted June 2020
Protocol Id: 2044
The Annexin V-FITC Early Apoptosis Detection Kit enables researchers to identify early apoptotic cells within a cell population. Annexin V-FITC conjugated protein binds to cell surfaces expressing phosphatidylserine, an early apotosis marker. Cells stained with propidium iodide (PI), a non-cell-permeable DNA dye, indicate necrotic cells. Cells stained with both PI and annexin V-FITC 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 charactersistic 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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