Annexin V (FITC)
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Price:
US$406.00
(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 (FITC) only and does not include 10X Binding Buffer, PI or 7-AAD reagents.
Target |
Annexin V |
Reactivity |
General (All species) |
Conjugation |
FITC |
Excitation/Emission |
499/515 |
Laser Line |
488 |
Form |
Liquid |
Storage |
Store at at 2-8°C in the dark. |
Validity |
12 months |
Buffer |
The exact formulation is proprietary. |
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: - 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.
- 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.
- Add 5 µl of Annexin V-FITC Reagent and 5 µl of DNA dye (PI, 7-AAD or DAPI) to each tube.
- Gently vortex the cells and incubate at room temperature for 15-20 min in the dark.
- 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.
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Research Articles on Annexin V