NBD-lipid Uptake Assay for Mammalian Cell Lines
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NBD-lipid Uptake Assay for Mammalian Cell Lines. / Herrera, Sara Abad; Grifell-Junyent, Marta; Pomorski, Thomas Günther.
In: Bio-protocol, Vol. 12, No. 4, 258649, 2022.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - NBD-lipid Uptake Assay for Mammalian Cell Lines
AU - Herrera, Sara Abad
AU - Grifell-Junyent, Marta
AU - Pomorski, Thomas Günther
N1 - Publisher Copyright: © 2022 The Authors.
PY - 2022
Y1 - 2022
N2 - All eukaryotic cells are equipped with transmembrane lipid transporters, which are key players in membrane lipid asymmetry, vesicular trafficking, and membrane fusion. The link between mutations in these transporters and disease in humans highlights their essential role in cell homeostasis. Yet, many key features of their activities, their substrate specificity, and their regulation remain to be elucidated. Here, we describe an optimized quantitative flow cytometry-based lipid uptake assay utilizing nitrobenzoxadiazolyl (NBD) fluorescent lipids to study lipid internalization in mammalian cell lines, which allows characterizing lipid transporter activities at the plasma membrane. This approach allows for a rapid analysis of large cell populations, thereby greatly reducing sampling variability. The protocol can be applied to study a wide range of mammalian cell lines, to test the impact of gene knockouts on lipid internalization at the plasma membrane, and to uncover the dynamics of lipid transport at the plasma membrane.
AB - All eukaryotic cells are equipped with transmembrane lipid transporters, which are key players in membrane lipid asymmetry, vesicular trafficking, and membrane fusion. The link between mutations in these transporters and disease in humans highlights their essential role in cell homeostasis. Yet, many key features of their activities, their substrate specificity, and their regulation remain to be elucidated. Here, we describe an optimized quantitative flow cytometry-based lipid uptake assay utilizing nitrobenzoxadiazolyl (NBD) fluorescent lipids to study lipid internalization in mammalian cell lines, which allows characterizing lipid transporter activities at the plasma membrane. This approach allows for a rapid analysis of large cell populations, thereby greatly reducing sampling variability. The protocol can be applied to study a wide range of mammalian cell lines, to test the impact of gene knockouts on lipid internalization at the plasma membrane, and to uncover the dynamics of lipid transport at the plasma membrane.
KW - Flow cytometry
KW - Lipid transport
KW - Mammalian cells
KW - NBD-lipid
KW - Plasma membrane
U2 - 10.21769/BioProtoc.4330
DO - 10.21769/BioProtoc.4330
M3 - Journal article
C2 - 35340299
AN - SCOPUS:85125915769
VL - 12
JO - Bio-protocol
JF - Bio-protocol
SN - 2331-8325
IS - 4
M1 - 258649
ER -
ID: 300911679