Description
Key facts
Detection method:Colorimetric/Fluorometric,
Sample types:Urine, Plasma, Tissue Extracts, Cell culture supernatant, Serum, Other biological fluids,
Assay type:Quantitative,
Assay time:1h,
Assay Platform:Microplate reader
Product details:
Cholesterol/ Cholesteryl Ester Assay Kit ab65359 provides a simple method for sensitive quantification of total cholesterol, free cholesterol, and cholesteryl esters, by colorimetric or fluorometric methods.
How the assay works
The majority of cholesterol in blood is in the form of cholesteryl esters. In the cholesterol assay protocol, these are hydrolyzed to cholesterol by cholesterol esterase. Cholesterol is then oxidized by cholesterol oxidase to yield hydrogen peroxide. Hydrogen peroxide is then processed by an enzyme causing a reaction with a probe to generate red color (λmax = 570 nm) and fluorescence (Ex/Em = 535/587 nm).
The assay detects total cholesterol (cholesterol and cholesteryl esters) when cholesterol esterase is included in the reaction, or free cholesterol when it is not included. The amount of cholesteryl ester can be determined by subtracting the value of free cholesterol from the total (cholesterol plus cholesteryl esters).
Cholesterol assay protocol summary
- Add samples and standards to wells.
- Add total cholesterol reaction mix (with esterase) or free cholesterol reaction mix (without esterase) and incubate for 60 min at 37°C.
- Analyze with microplate reader.
Related Cholesterol assay products
Cholesterol Assay Kit - HDL and LDL/VLDL
ab65390
is identical to Cholesterol/ Cholesteryl Ester Assay Kit ab65359, except for the addition of a buffer and protocol for separation of HDL and LDL/VLDL fractions.
Related and recommended products
Other cholesterol assay kits include:
- Cell-based Cholesterol assay kit
ab133116
- Cholesterol Efflux assay kit
ab196985
- Cholesterol Uptake assay kit
ab236212
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Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K603 Total Cholesterol and Cholesteryl Ester Colorimetric/Fluorometric Assay Kit. K603-100 is the same size as the 100 test size of ab65359.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Cholesterol/cholesteryl ester (CE) plays a mechanical role in the body by transporting lipids. Known also as 'cholesteryl esters' these molecules consist of cholesterol bound to long-chain fatty acids. CE exhibits a molecular mass that varies depending on the fatty acid chain length. These molecules are typically found within plasma and intracellular lipid droplets in tissues such as the liver and intestines. They mediate cholesterol transport and storage maintaining cellular lipid homeostasis.
Biological function summary
Cholesteryl esters facilitate cholesterol influx and efflux by serving as precursor molecules for various hormones and vitamin D. They do not exist in isolation but rather as part of larger lipid complexes including chylomicrons very-low-density lipoproteins (VLDL) and low-density lipoproteins (LDL). These lipid complexes play an essential role in delivering cholesterol to peripheral tissues and returning excess to the liver for excretion or reprocessing.
Pathways
Cholesteryl esters are involved in lipid metabolism processes such as the reverse cholesterol transport and the esterification of cholesterol through the action of lecithin-cholesterol acyltransferase (LCAT). This esterification process converts free cholesterol into cholesteryl esters facilitating their transport within lipoproteins. High-density lipoprotein (HDL) serves as a pathway partner circulating cholesteryl esters back to the liver where they undergo hydrolysis and re-utilization.
Cholesteryl esters are significant in the development of atherosclerosis and coronary artery disease. Dysregulation of CE metabolism can lead to the accumulation of lipid-rich plaques in blood vessel walls catalyzed by proteins like apolipoproteins such as ApoB-100 and ApoE. Excessive LDL and CE deposition in arterial walls are associated with plaque formation culminating in cardiovascular diseases. Understanding the role of these compounds and their associated pathways is important for developing treatments against lipid disorders.
