Description
Key facts
Detection method:Fluorescent,
Sample types:Adherent cells,
Assay type:Cell-based,
Assay time:1h 15m,
Assay Platform:Fluorescence microscope
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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Cholesterol sometimes referred to as fchol is a lipid molecule with a molecular mass of approximately 386.65 Daltons. It is an essential component of cell membranes and is found in the plasma membranes of cells throughout the body. Cholesterol primarily resides in the membranes where it modulates fluidity and mechanical properties. Its expression occurs in various tissues but it is abundant in the liver which plays an important role in its synthesis and regulation.
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.
Cholesterol is closely associated with atherosclerosis and cardiovascular disease. Elevated levels of low-density lipoprotein (LDL) cholesterol contribute to plaque formation in arteries increasing the risk of heart attack and stroke. On the other hand high-density lipoprotein (HDL) cholesterol helps remove excess cholesterol from tissues including the arterial walls. Statins a class of drugs that lower LDL cholesterol levels target HMG-CoA reductase and are commonly used in treating hypercholesterolemia and cardiovascular diseases.
