Key features and details
- Assay type: Cell-based (quantitative)
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Adherent cells, Purified protein
Product nameCholesterol Efflux Assay Kit (Cell-based)
Sample typeAdherent cells, Purified protein
Assay typeCell-based (quantitative)
Species reactivityReacts with: Mammals, Other species
Cholesterol Efflux Assay Kit ab196985 is a high-throughput, cell-based screening assay for measuring cholesterol efflux. The cholesterol efflux assay protocol uses fluorescently-labeled cholesterol (Ex/Em 482/515 nm). It provides a safe, sensitive and reproducible method for measuring cholesterol efflux.
This cholesterol efflux assay can be used to:
- screen small molecules for their effect on cholesterol efflux as a part of drug discovery program.
Cholesterol efflux assay protocol summary:
- label cells with labeling and equilibration mix overnight
- wash cells, treat as required and incubate
- transfer cell supernatant to microplate, and separately solubilize cells with cell lysis buffer
- analyze supernatant and cell lysates with microplate reader
This product is manufactured by BioVision, an Abcam company and was previously called K582 Cholesterol Efflux Fluorometric Assay Kit (cell-based). K582-100 is the same size as the 100 test size of ab196985.
Cholesterol efflux from the peripheral tissues and cells in atherosclerotic plaque is an initial and critical step in Reverse Cholesterol Transport (RCT). RCT is the process by which extra hepatic cells, including macrophage-derived foam cells in arterial atherosclerotic plaque, transport excessive cholesterol back to the liver for bile acid synthesis and excretion, thus lowering the peripheral lipid burden.
Other cholesterol assay kits include:
- HDL and LDL/VLDL Cholesterol assay kit ab65390
- Cell-based Cholesterol assay kit ab133116
- Cholesterol/Cholesterol Ester assay kit ab65359
- Cholesterol Uptake assay kit ab236212
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 100 tests Lysis Buffer I 1 x 20ml Equilibration Buffer 1 x 5ml Labeling Reagent 1 x 5ml Cholesterol Uptake Positive Control 1 x 1ml Reagent A 1 x 10µl Reagent B 1 x 50µl Serum Treatment Reagent 1 x 1ml
Percentage (%) Labeled Cholesterol Efflux. J774.1 cells were labeled with the fluorescent cholesterol Labeling Media and treated with various cholesterol acceptors such as 2% LDL/VLDL-depleted Human Serum (2 µl/well), purified human HDL (25 µg/ml, 2.5 µg/well) or Positive Control reagent. Cholesterol efflux is expressed as percentage of fluorescently-labeled cholesterol transferred from labeled adherent macrophages to designated cholesterol acceptor present in supernatant over a 4 hour time period, with no treatment control (efflux to medium containing no acceptor) percentage subtracted (mean ± SD of N = 5-6 wells).
ab196985 has been referenced in 30 publications.
- Katakam S et al. Necrotic debris and STING exert therapeutically relevant effects on tumor cholesterol homeostasis. Life Sci Alliance 5:N/A (2022). PubMed: 34983824
- Sauter M et al. Apolipoprotein E derived from CD11c+ cells ameliorates atherosclerosis. iScience 25:103677 (2022). PubMed: 35036868
- Balamurugan K et al. PHLPP1 promotes neutral lipid accumulation through AMPK/ChREBP-dependent lipid uptake and fatty acid synthesis pathways. iScience 25:103766 (2022). PubMed: 35141506
- Huwait E et al. Thymoquinone (TQ) Inhibits Inflammation and Migration of THP-1 Macrophages: Mechanistic Insights into the Prevention of Atherosclerosis Using In-Vitro and In-Silico Analysis. Curr Issues Mol Biol 44:1740-1753 (2022). PubMed: 35723378
- Wang M et al. Qihuang Zhuyu Formula Attenuates Atherosclerosis via Targeting PPARγ to Regulate Cholesterol Efflux and Endothelial Cell Inflammation. Oxid Med Cell Longev 2022:2226168 (2022). PubMed: 36518993