Visualizing Loss of Plasma Membrane Lipid Asymmetry Using Annexin V Staining
Research output: Contribution to journal › Journal article › Research › peer-review
Loss of plasma membrane lipid asymmetry contributes to many cellular functions and responses, including apoptosis, blood coagulation, and cell fusion. In this protocol, we describe the use of fluorescently labeled annexin V to detect loss of lipid asymmetry in the plasma membrane of adherent living cells by fluorescence microscopy. The approach provides a simple, sensitive, and reproducible method to detect changes in lipid asymmetry but is limited by low sample throughput. The protocol can also be adapted to other fluorescently labeled lipid-binding proteins or peptide probes. To validate the lipid binding properties of such probes, we additionally describe here the preparation and use of giant unilamellar vesicles as simple model membrane systems that have a size comparable to cells.
Original language | English |
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Article number | e4754 |
Journal | Bio-protocol |
Volume | 13 |
Issue number | 14 |
ISSN | 2331-8325 |
DOIs | |
Publication status | Published - 2023 |
Bibliographical note
Publisher Copyright:
© 2023 The Authors; exclusive licensee Bio-protocol LLC.
- Confocal microscopy, Giant vesicle, Lipid asymmetry, Lipid-binding protein, Mammalian cells, Plasma membrane
Research areas
ID: 364546282