Annexin V (AF780)

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Catalogue No: abx092340
Price: US$232.00
(Size: 100 tests)

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Annexin V (AF780)

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Datasheet SDS
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 (AF780) only and does not include 10X Binding Buffer, PI or 7-AAD reagents.

Target Annexin V
Research Area Cell Death, Apoptosis
Reactivity General (All species)
Tested Applications FCM
Conjugation AF780
Form Liquid
Storage Store at 2-8°C in the dark.
Shelf Life 12 months.
Availability Shipped within 5-10 working days.
Note THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
Directions for use Suggested Staining Procedure:
  1. Induce apoptosis of suspension cells with reagents of interest. Collect cell cultures, centrifuge at 300 × g for 5 min and discard the supernatant. Add PBS to wash the cells and resuspend the cells gently. Carry out cell counting.
  2. Split the cell suspension into tubes, each containing 1-5 × 105 cells. Centrifuge at 300 × g for 5 min and discard the supernatant. Add PBS to wash the cells, then discard the supernatant. Add 500 µl of 1X Annexin V Binding Buffer (abx090621) to resuspend the cells.
  3. Add 5 µl of Annexin V (AF780) Reagent and 5 µl of DNA dye (PI, 7-AAD or DAPI) to each tube.
  4. Gently vortex the cells and incubate at room temperature for 15-20 min in the dark.
  5. Analyze the cells immediately with appropriate machine settings, or place the cells on ice in the dark and analyze within 1 h.
Notes:
  • When detecting adherent cells, the suspension cells generated after induction of apoptosis should be collected and detected together with the subsequently collected adherent cells.
  • Mechanical damage caused by digestion of adherent cells should be avoided. The trypsin digestion solution should not contain EDTA, because EDTA will affect the binding of Annexin V to phosphatidylserine. If trypsin-containing EDTA is used, cells should be washed thoroughly after harvesting to ensure that EDTA is removed.
  • Analyze as soon as possible after staining. Avoid extended exposure of the samples to direct light to protect the fluorophores from quenching.
  • Wear appropriate personal protective equipment (PPE), such as lab coats, safety glasses, and disposable gloves, when carrying out the assay.
Research Articles on Annexin V


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