Invitrogen eBioscience Annexin V Apoptosis Detection Kit eFluor 450
Compatible with intracellular staining. Pre-titrated and tested on mouse thymocytes cultured overnight in medium (to induce apoptosis) or on Jurkat cells treated with 10 µM Camptothecin for 4 hours. Both the Annexin V eFluor 450 and the 7-AAD components can be used at 5µL per test.
Manufacturer: Invitrogen 88800672
The components in this kit have been pre-titrated and tested on mouse thymocytes cultured overnight in medium (to induce apoptosis) or on Jurkat cells treated with 10µM Camptothecin for 4 hours. Both the Annexin V eFluor 450 and the 7-AAD components can be used at 5µL per test and should be protected from light.
Annexins are a family of calcium-dependent phospholipid-binding proteins that preferentially bind phosphatidylserine (PS). Under normal physiologic conditions, PS is predominantly located in the inner leaflet of the plasma membrane. Upon initiation of apoptosis, PS loses its asymmetric distribution across the phospholipid bilayer and is translocated to the extracellular membrane leaflet marking cells as targets of phagocytosis. Once on the outer surface of the membrane, PS can be detected by fluorescently labeled Annexin V in a calcium-dependent manner.
In early-stage apoptosis, the plasma membrane excludes viability dyes such as propidium iodide (PI), 7-AAD, or Fixable Viability Dyes such as eFluor 660 or eFluor 780. These cells will stain with Annexin V but not a viability dye, thus distinguishing cells in early apoptosis. However, in late stage apoptosis, the cell membrane loses integrity thereby allowing Annexin V to also access PS in the interior of the cell. A viability dye can be used to resolve these late-stage apoptotic and necrotic cells (Annexin V, viability dye-positive) from the early-stage apoptotic cells (Annexin V positive, viability dye-negative).
|Fluorescence Microscope, Flow Cytometer|
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