(Size: 100 tests)
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Annexin V is used as a probe to detect cells that have expressed phosphatidylserine (PS) on the cell surface, an event found in apoptosis as well as other forms of cell death. The Annexin V affinity assay typically uses a conjugate of annexin V and a fluorescent or enzymatic label, biotin or other tags, or a radioelement, in a suitable buffer (Annexin V binding to PS is Ca2+
dependent). The assay combines Annexin V staining of PS membrane events with the staining of the cell nucleus with PI or AAD-7 to distinguish living cells from dead cells. Annexin V apoptosis detection is based on the observation that soon after initiating apoptosis, cells translocate the membrane phosphatidylserine (PS) from the inner (cytoplasmic-facing) leaflet of the plasma membrane to the cell surface. Once on the cell surface, PS can be easily detected by staining with a fluorescent conjugate of Annexin V, a protein that has a high affinity for PS. Detection can be analyzed by flow cytometry or by fluorescence microscopy.
This product contains Annexin V (APC) only and does not include 10X Binding Buffer, PI or 7-AAD reagents.
Research Articles on Annexin V
||General (All species)
||Store undiluted at 4 °C. Avoid exposure to light. Do not freeze.
||PBS, pH 7.2, 0.09% NaN3.
|Directions for use
- Dilute the 10X Binding Buffer solution to 1X Working Binding Buffer solution with distilled water.
- Harvest cells (about 1 × 105 cells per test), then wash once with cold PBS. Remove the PBS from the cell pellet.
- Wash again with cold 1X Working Binding Buffer, then centrifuge at 300 × g for 10 min at room temperature. Remove the Binding Buffer from the cell pellet.
- Resuspend cells in cold 1X Working Binding Buffer to a concentration of 1 × 106 cells/ml.
- Add 100 µl of cells (1 × 105 cells) to each appropriate tube.
- Add 5 µl of Annexin V-APC to the appropriate tubes.
- Gently vortex each tube and incubate for 10 minutes at room temperature in dark.
- Add 5 µl of PI or 7-AAD solution and incubate for 5 min at room temperature in dark.
- Wash cells once in PBS, then resuspend in PBS.
- Analyze by flow cytometry within 4 hours.
||Shipped within 5-10 working days.
||This product is for research use only.