Visualizing Loss of Plasma Membrane Lipid Asymmetry Using Annexin V Staining
Research output: Contribution to journal › Journal article › Research › peer-review
Standard
Visualizing Loss of Plasma Membrane Lipid Asymmetry Using Annexin V Staining. / Baum, Julia F.; Uzun, Huriye D.; Pomorski, Thomas Günther.
In: Bio-protocol, Vol. 13, No. 14, e4754, 2023.Research output: Contribution to journal › Journal article › Research › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Visualizing Loss of Plasma Membrane Lipid Asymmetry Using Annexin V Staining
AU - Baum, Julia F.
AU - Uzun, Huriye D.
AU - Pomorski, Thomas Günther
N1 - Publisher Copyright: © 2023 The Authors; exclusive licensee Bio-protocol LLC.
PY - 2023
Y1 - 2023
N2 - 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.
AB - 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.
KW - Confocal microscopy
KW - Giant vesicle
KW - Lipid asymmetry
KW - Lipid-binding protein
KW - Mammalian cells
KW - Plasma membrane
U2 - 10.21769/BioProtoc.4754
DO - 10.21769/BioProtoc.4754
M3 - Journal article
C2 - 37497452
AN - SCOPUS:85166533756
VL - 13
JO - Bio-protocol
JF - Bio-protocol
SN - 2331-8325
IS - 14
M1 - e4754
ER -
ID: 364546282