Visualizing Loss of Plasma Membrane Lipid Asymmetry Using Annexin V Staining
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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.
Originalsprog | Engelsk |
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Artikelnummer | e4754 |
Tidsskrift | Bio-protocol |
Vol/bind | 13 |
Udgave nummer | 14 |
ISSN | 2331-8325 |
DOI | |
Status | Udgivet - 2023 |
Bibliografisk note
Funding Information:
We gratefully acknowledge Michelle Werner for technical assistance. This protocol was adapted from our previous work (Weingärtner et al., 2012; Grifell-Junyent et al., 2022; Herrera et al., 2022). The work was supported by the Lundbeckfonden (R221-2016-1005 to T.G.P.) and an instrument grant from the Deutsche Forschungsgemeinschaft (INST 213/886-1 FUGG to T.G.P.). HDU is a scholar of the Friedrich Ebert Foundation.
Publisher Copyright:
© 2023 The Authors; exclusive licensee Bio-protocol LLC.
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