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| Description |
The Annexin V MicroBead Kit was developed for positive selection or depletion of cells based on their ability to bind Annexin V.
In most normal, viable eukaryotic cells the negatively charged phospholipid phosphatidylserine (PS) is located in the cytosolic leaflet of the plasma membrane lipid bilayer. PS redistribution from the inner to the outer leaflet is an early and widespread event during apoptosis. However, in necrosis, PS becomes accessible due to the disruption of membrane integrity. Apart from necrosis and apoptosis, PS also becomes accessible in activated platelets, in certain cell anomalies such as sickle cell anaemia1, in erythrocyte senescence, upon degranulation of mast cells, and in certain stages of B cell differentiation. PS exposure also serves as a trigger for the recognition and removal of cells by macrophages.
Annexin V is a 35 kDa phospholipid-binding protein and a major cell membrane component of macrophages and other phagocytic cell types. The Annexin V MicroBead Kit includes Annexin V MicroBeads and Annexin V Binding Buffer. |
| Applications |
| Annexin V has a high affinity to PS in the presence of physiological concentrations of calcium (Ca2+). Therefore, the Annexin V MicroBead Kit is used for the isolation of cells with exposed PS, including sickle cells and apoptotic cells.1–3 The Annexin V MicroBead Kit has been further applied to the purification of apoptotic and non-apoptotic neutrophils to study the intracellular signaling pathways that regulate neutrophil survival.4 In a study on the roles of different cytosolic phospholipase A2s in apoptosis, the Annexin V MicroBead Kit has been used for sensitive assessment of PS externalization.5 |
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| Figure 1 |
| Isolation of PS-exposing cells using the Annexin V MicroBead Kit. |
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| Figure 2 |
| Enrichment of dead (apoptotic/necrotic) cells using the Annexin V MicroBead Kit. Murine thymocytes were treated with CD95 (anti-Fas) monoclonal antibody for 3 hours at 37 °C. PS-exposing cells were enriched by labeling with Annexin V MicroBeads followed by separation on an MS Column in the magnetic field of a MiniMACS™ Separator. Cell fractions were stained with Annexin V-FITC. |
| A: Murine thymocytes after treatment with CD95 (anti-Fas) antibody |
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| B: Positive fraction enriched for PS-exposing cells |
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| MACS References |
| 1. Kuypers et al. (1996) Blood 87: 1179–1187.[181] |
| 2. Rickers et al. (1998) Eur. J. Immunol. 28: 296–304.[425] |
| 3. Dressel et al. (2000) J. Immuno. 164: 2362–2371.[572] |
| 4. Khwaja and Tatton (1999) Blood 94: 291–301.[673] |
| 5. Atsumi et al. (2000) J. Biol. Chem. 16: 18248–18258.[969] |
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