Cholesterol Assay Kit (Cell-Based)
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(52 Publications)
- Fluorescence Microscopy
Supplier Data
Fluorescence Microscopy - Cholesterol Assay Kit (Cell-Based) (AB133116)
Accumulation of cholesterol inside HepG2 cells in response to 1.25 µM U-18666A.
HepG2 cells were seeded in a 96-well plate at a density of 3 x 104 cells/well and cultured overnight. The next day, cells were treated with DMSO (vehicle) or 1.25 µM U-18666A for 48 hours. Panel A : Cells treated with DMSO alone demonstrate that majority of cholesterol is localized on the plasma membrane. Panel B : U-18666A treatment for 48 hours induces intracellular accumulation.
- Fluorescence Microscopy
Lab
Fluorescence Microscopy - Cholesterol Assay Kit (Cell-Based) (AB133116)
HepG2 cells in response to 2 μM U-18666A.
HepG2 cells were seeded in a 96-well plate at a density of 2.4 x 104 cells/well and cultured overnight. The next day, cells were treated with 2 μM U-18666A for 48 hours. U-18666A treatment for 48 hours induces intracellular accumulation.
- Fluorescence Microscopy
Lab
Fluorescence Microscopy - Cholesterol Assay Kit (Cell-Based) (AB133116)
HepG2 cells in response to 1 μM U-18666A.
HepG2 cells were seeded in a 96-well plate at a density of 2.4 x 104 cells/well and cultured overnight. The next day, cells were treated with 1 μM U-18666A for 48 hours. U-18666A treatment for 48 hours induces intracellular accumulation.
- Fluorescence Microscopy
Lab
Fluorescence Microscopy - Cholesterol Assay Kit (Cell-Based) (AB133116)
Accumulation of cholesterol inside HepG2 cells - Control.
HepG2 cells were seeded in a 96-well plate at a density of 2.4 x 104 cells/well and cultured overnight. The next day, cells were treated with DMSO (vehicle) for 48 hours. Cells demonstrate that majority of cholesterol is localized on the plasma membrane.
- Fluorescence Microscopy
Lab
Fluorescence Microscopy - Cholesterol Assay Kit (Cell-Based) (AB133116)
HepG2 cells in response to 0.25 μM U-18666A.
HepG2 cells were seeded in a 96-well plate at a density of 2.4 x 104 cells/well and cultured overnight. The next day, cells were treated with 0.25 μM U-18666A for 48 hours. U-18666A treatment for 48 hours induces intracellular accumulation.
- Fluorescence Microscopy
Lab
Fluorescence Microscopy - Cholesterol Assay Kit (Cell-Based) (AB133116)
HepG2 cells in response to 0.5 μM U-18666A.
HepG2 cells were seeded in a 96-well plate at a density of 2.4 x 104 cells/well and cultured overnight. The next day, cells were treated with 0.5 μM U-18666A for 48 hours. U-18666A treatment for 48 hours induces intracellular accumulation.
Complementary Products
Product details
Cholesterol Assay Kit (Cell-Based) (ab133116) includes filipin III, fixative, and wash buffer in a ready to use format. It provides a simple fluorometric method to study mechanisms and biological factors that regulate cholesterol metabolism or movement within cells. A cholesterol trafficking inhibitor, U-18666A, is included as a positive control.
Cholesterol is both an important structural component of cell membranes and an early intermediate in hormone and bile acid biosynthesis. Cholesterol is not uniformly distributed among cellular membranes / organelle membranes, but rather there are structurally and kinetically distinct cholesterol rich and poor domains.
How the cell-based cholesterol assay works
Cholesterol Assay Kit (Cell-Based) includes filipin III, fixative, and wash buffer in a ready to use format.
Filipin III is the predominant isomer of filipin, the collective name given to four isomeric polyene macrolides isolated from cultures of S.filipinensis.
Filipin has been widely used as a probe for sterol location in biological membranes. Interaction with cholesterol alters the filipin absorption and fluorescence spectra allowing visualization with a fluorescence microscope capable of excitation at 340-380 nm and emission at 385-470 nm.
Filipin’s ease of use makes it a convenient tool for the histochemical identification of unesterified cholesterol both in vitro and in vivo.
Cholesterol assay protocol summary:
- remove medium from cells
- fix with fixative solution for 10 min
- wash with wash buffer 3 times for 5 min
- add Filipin III solution and incubate for 30-60 min
- wash twice for 5 min
- analyze with fluorescent microscope
Other cholesterol assay kits include:
- HDL and LDL/VLDL Cholesterol assay kit ab65390
- Cholesterol/Cholesterol Ester assay kit ab65359
- Cholesterol Efflux assay kit ab196985
- Cholesterol Uptake assay kit ab236212
REACH authorisation
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.
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Properties and storage information
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Storage information
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Cholesterol is important in maintaining the structural integrity and organization of membranes. It is also part of lipid rafts which are specialized microdomains within the cell membrane that facilitate cellular signaling. Cholesterol interacts with proteins and receptors embedded in the membrane influencing their function. It does not form a complex itself but affects complexes within the membrane like lipid rafts and signaling platforms.
Pathways
Cholesterol is a critical component of the steroid biosynthesis pathway and the bile acid synthesis pathway. It serves as a precursor for steroid hormones including cortisol and sex hormones as well as bile acids which are essential for digestion and lipid absorption. Proteins such as HMG-CoA reductase and the low-density lipoprotein receptor (LDLR) are key players related to cholesterol in these pathways regulating its synthesis and uptake in cells.
Product protocols
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Publications (52)
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Science advances 11:eadr5703 PubMed40333988
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eLife 13: PubMed40270220
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eLife 13: PubMed40184172
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Nature communications 16:2278 PubMed40055353
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Molecular neurodegeneration 20:15 PubMed39901180
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eLife 12: PubMed39713930
2024
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Frontiers in endocrinology 15:1383448 PubMed39544235
2024
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Cell 187:5336-5356.e30 PubMed39137777
2024
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Molecular metabolism 82:101913 PubMed38458567
2024
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Cell stem cell 31:359-377.e10 PubMed38458178
2024
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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