NBD-lipid Uptake Assay for Mammalian Cell Lines

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

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

Harvard

Herrera, SA, Grifell-Junyent, M & Pomorski, TG 2022, 'NBD-lipid Uptake Assay for Mammalian Cell Lines', Bio-protocol, vol. 12, no. 4, 258649. https://doi.org/10.21769/BioProtoc.4330

APA

Herrera, S. A., Grifell-Junyent, M., & Pomorski, T. G. (2022). NBD-lipid Uptake Assay for Mammalian Cell Lines. Bio-protocol, 12(4), [258649]. https://doi.org/10.21769/BioProtoc.4330

Vancouver

Herrera SA, Grifell-Junyent M, Pomorski TG. NBD-lipid Uptake Assay for Mammalian Cell Lines. Bio-protocol. 2022;12(4). 258649. https://doi.org/10.21769/BioProtoc.4330

Author

Herrera, Sara Abad ; Grifell-Junyent, Marta ; Pomorski, Thomas Günther. / NBD-lipid Uptake Assay for Mammalian Cell Lines. In: Bio-protocol. 2022 ; Vol. 12, No. 4.

Bibtex

@article{74efa34d77884200b1bf5429f580ff64,
title = "NBD-lipid Uptake Assay for Mammalian Cell Lines",
abstract = "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. ",
keywords = "Flow cytometry, Lipid transport, Mammalian cells, NBD-lipid, Plasma membrane",
author = "Herrera, {Sara Abad} and Marta Grifell-Junyent and Pomorski, {Thomas G{\"u}nther}",
note = "Publisher Copyright: {\textcopyright} 2022 The Authors.",
year = "2022",
doi = "10.21769/BioProtoc.4330",
language = "English",
volume = "12",
journal = "Bio-protocol",
issn = "2331-8325",
publisher = "bio-protocol",
number = "4",

}

RIS

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