YILIMART - Annexin V-FITC Kit

This kit is suitable for the identification and enumeration of dead cells, such as apoptotic or necrotic cells, by flow cytometry.

Background information

The Annexin V-FITC Kit contains reagents for the simultaneous fluorescent detection of apoptotic, necrotic, and dead cells through the specific binding of FITC-labeled Annexin V to phosphotidylserine (PS). The kit consists of Annexin V-FITC, Annexin V Binding Buffer, and Propidium Iodide Solution.

In most normal, viable eukaryotic cells, the negatively charged phospholipid PS is located in the cytosolic leaflet of the plasma membrane lipid bilayer.

¹

PS redistribution from the inner to the outer leaflet is an early and widespread event during apoptosis.

^1,2

However, in necrosis, PS becomes accessible due to the disruption of membrane integrity.

²

Apart from necrosis and apoptosis, PS also becomes accessible in activated platelets

³

, in certain cell anomalies such as sickle cell anemia

⁴

, in erythrocyte senescence

⁵

, upon degranulation of mast cells

⁶

, and in certain stages of B cell differentiation

⁷

. PS exposure by apoptotic cells serves as a trigger for the recognition and removal by macrophages.

^8,9

Annexin V is a 35 kDa phospholipid-binding protein and has a high affinity for PS in the presence of physiological concentrations of calcium (Ca

²⁺

).

¹⁰

Figure 1
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Figure 1

Jurkat cells cultured with staurosporine (50 nM) for 15 h, were stained with Annexin V-FITC and PI and analyzed by flow cytometry.

Jurkat cells cultured with staurosporine (50 nM) for 15 h, were stained with Annexin V-FITC and PI and analyzed by flow cytometry.
Figure 2
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Figure 2

Staining principle of PS-exposing cells with Annexin V-FITC.

Staining principle of PS-exposing cells with Annexin V-FITC.

Selected references
1. Koopman, G. et al. (1994) Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84: 1415-1420
2. Martin, S. J. et al. (1995) Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J. Exp. Med. 182: 1545-1556
3. Thiagarajan, P. et al. (1990) Binding of annexin V/placental anticoagulant protein I to platelets. Evidence for phosphatidylserine exposure in the procoagulant response of activated platelets. J. Biol. Chem. 265: 17420-17423
4. Kuypers, F. A. et al. (1996) Detection of altered membrane phospholipid asymmetry in subpopulations of human red blood cells using fluorescently labeled annexin V. Blood 87: 1179-1187
5. Schroit, A. J. et al. (1991) Transbilayer movement of phospholipids in red cell and platelet membranes. Biochim. Biophys. Acta 1071: 313-329
6. Demo et al. (1999) Quantitative measurement of mast cell degranulation using a novel flow cytometric annexin-V binding assay. Cytometry 36: 340-348
7. Dillon, S. R. et al. (2001) Annexin V binds to positively selected B cells. J. Immunol. 166: 58-71
8. Fadok, V. A. et al. (1992) Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J. Immunol. 148: 2207-2216
9. Fadok, V. A. et al. (2000) A receptor for phosphatidylserine-specific clearance of apoptotic cells. Nature 405: 85-90
10. Moss, S. E. et al. (1991) Diversity in the Annexin family. In: Novel Calcium Binding Proteins, Springer Verlag : 535-566

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Annexin V-FITC Kit

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Background information

Figure 1

Figure 1

Figure 2

Figure 2

Selected references

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