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A High-throughput, Fluorescence-based Cholesterol Efflux Assay
EP27367
Poster Title: A High-throughput, Fluorescence-based Cholesterol Efflux Assay
Submitted on 15 Mar 2018
Author(s): Sonika Saddar, Tasnim Kothambawala, Nick Chacos, Preeti Saini, Grigoriy Tchaga and Gordon Yan
Affiliations: BioVision Inc.
Poster Views: 737
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Poster Information
Abstract: Numerous studies have established a negative correlation between
cellular cholesterol efflux and cardiovascular disease. Cholesterol efflux
from peripheral tissues and cells by a process called Reverse Cholesterol
Transport (RCT). RCT allows extrahepatic cells, including macrophagederived
foam cells in arterial atherosclerotic plaque, to transport
excessive cholesterol back to the liver for bile acid synthesis and
excretion, thus lowering the peripheral lipid burden. Standard protocols
for quantitation of cholesterol efflux involve labeling cells with tritiated
cholesterol and measuring the release of labeled sterol in a 12 or 24 well
plate. This protocol, however is not ideal for high-throughput screening of
large numbers of serum samples or screening compounds/small
molecules for their effect on cholesterol efflux. We report here a simple,
convenient and high-throughput cell-based assay using fluorescentlylabeled
cholesterol in a 96-well plate format. Our results demonstrate that
the percentage cholesterol efflux elicited by 2% ApoB-depleted normal
human serum in J774 macrophage cells is comparable to those
published in literature. Our assay method utilizes a positive control for
this assay which induces efflux independent of any receptor, that is used
as a measure of the reliability of the assay. This assay is highly
reproducible with low inter-assay variability (≤ 10%). We demonstrate that
this is an efficient method of measuring cholesterol efflux in a cell-based
assay. The increased sensitivity of the assay coupled with its highthroughput
screening capability enables scientists to screen large
libraries of reagents and serum samples and provides a valuable tool for
drug discovery in the field of cardiovascular disease research.
Summary: • BioVision’s Cholesterol Efflux Assay Kit is a high-throughput screening assay
for measuring cholesterol efflux in cells using fluorescently-labelled
cholesterol.
• Cholesterol Efflux Assay provides a safe, sensitive and reproducible method
of measuring cholesterol efflux.
• Several factors that modifies Reverse Cholesterol Transport and cholesterol
efflux can be tested using this kit.
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