An assay to evaluate the capacity of cholesterol acceptors using BODIPY-cholesterol in cells

Ana Caroline Raulin; Chia Chen Liu; Guojun Bu

doi:10.1016/j.xpro.2022.101976

An assay to evaluate the capacity of cholesterol acceptors using BODIPY-cholesterol in cells

Ana Caroline Raulin, Chia Chen Liu, Guojun Bu

Neuroscience

Research output: Contribution to journal › Article › peer-review

Abstract

Cholesterol is a structural component of cell membranes. Most cells are incapable of its catabolism, and intracellular cholesterol accumulation is linked to several disorders including cardiovascular and neurodegenerative diseases. Cholesterol efflux, essential to its metabolism, is dependent on acceptors such as apolipoproteins. Here, we describe an assay to evaluate the capacity of cholesterol acceptors. Cells are treated with an analog of cholesterol tagged with fluorescent BODIPY. Addition of an acceptor leads to BODIPY-cholesterol efflux, measured using a plate reader. For complete details on the use and execution of this protocol, please refer to Liu et al. (2021).¹

Original language	English (US)
Article number	101976
Journal	STAR Protocols
Volume	4
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2022.101976
State	Published - Mar 17 2023

Keywords

Cell-based Assays
Molecular/Chemical Probes

ASJC Scopus subject areas

Neuroscience(all)
Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)

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10.1016/j.xpro.2022.101976

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title = "An assay to evaluate the capacity of cholesterol acceptors using BODIPY-cholesterol in cells",

abstract = "Cholesterol is a structural component of cell membranes. Most cells are incapable of its catabolism, and intracellular cholesterol accumulation is linked to several disorders including cardiovascular and neurodegenerative diseases. Cholesterol efflux, essential to its metabolism, is dependent on acceptors such as apolipoproteins. Here, we describe an assay to evaluate the capacity of cholesterol acceptors. Cells are treated with an analog of cholesterol tagged with fluorescent BODIPY. Addition of an acceptor leads to BODIPY-cholesterol efflux, measured using a plate reader. For complete details on the use and execution of this protocol, please refer to Liu et al. (2021).1",

keywords = "Cell-based Assays, Molecular/Chemical Probes",

author = "Raulin, {Ana Caroline} and Liu, {Chia Chen} and Guojun Bu",

note = "Funding Information: The authors thank Dr. Na Zhao and Dr. Yuka Martens for technical support. This research was supported by the National Institute of Health (NIH) (grants U19AG069701 to C.-C.L. and G.B.; R01AG062110 to C.-C.L.), a Cure Alzheimer{\textquoteright}s Fund (G.B.), and a grant from Brightfocus (C.-C.L.), and A-C.R. is supported by a BrightFocus Fellowship. Publisher Copyright: {\textcopyright} 2022 The Authors",

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doi = "10.1016/j.xpro.2022.101976",

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AU - Raulin, Ana Caroline

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AU - Bu, Guojun

N1 - Funding Information: The authors thank Dr. Na Zhao and Dr. Yuka Martens for technical support. This research was supported by the National Institute of Health (NIH) (grants U19AG069701 to C.-C.L. and G.B.; R01AG062110 to C.-C.L.), a Cure Alzheimer’s Fund (G.B.), and a grant from Brightfocus (C.-C.L.), and A-C.R. is supported by a BrightFocus Fellowship. Publisher Copyright: © 2022 The Authors

PY - 2023/3/17

Y1 - 2023/3/17

N2 - Cholesterol is a structural component of cell membranes. Most cells are incapable of its catabolism, and intracellular cholesterol accumulation is linked to several disorders including cardiovascular and neurodegenerative diseases. Cholesterol efflux, essential to its metabolism, is dependent on acceptors such as apolipoproteins. Here, we describe an assay to evaluate the capacity of cholesterol acceptors. Cells are treated with an analog of cholesterol tagged with fluorescent BODIPY. Addition of an acceptor leads to BODIPY-cholesterol efflux, measured using a plate reader. For complete details on the use and execution of this protocol, please refer to Liu et al. (2021).1

AB - Cholesterol is a structural component of cell membranes. Most cells are incapable of its catabolism, and intracellular cholesterol accumulation is linked to several disorders including cardiovascular and neurodegenerative diseases. Cholesterol efflux, essential to its metabolism, is dependent on acceptors such as apolipoproteins. Here, we describe an assay to evaluate the capacity of cholesterol acceptors. Cells are treated with an analog of cholesterol tagged with fluorescent BODIPY. Addition of an acceptor leads to BODIPY-cholesterol efflux, measured using a plate reader. For complete details on the use and execution of this protocol, please refer to Liu et al. (2021).1

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KW - Molecular/Chemical Probes

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An assay to evaluate the capacity of cholesterol acceptors using BODIPY-cholesterol in cells

Abstract

Keywords

ASJC Scopus subject areas

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