Cholesterol/ Cholesteryl Ester Assay Kit ab65359 provides a simple method for sensitive quantification of total cholesterol, free cholesterol, and cholesteryl esters, by colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) plate reader.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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Publications
- Translational psychiatry 14:132024Olanzapine, risperidone and ziprasidone differently affect lysosomal function and autophagy, reflecting their different metabolic risk in patients.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMarco Pozzi,Chiara Vantaggiato,Francesca Brivio,Genny Orso,Maria Teresa BassiPubMed 38191558
- Communications biology 7:82024Cholesterol modulates type I/II TGF-β receptor complexes and alters the balance between Smad and Akt signaling in hepatocytes.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRoohi Chaudhary,Laureen S Goodman,Sai Wang,Anastasia Asimakopoulos,Ralf Weiskirchen,Steven Dooley,Marcelo Ehrlich,Yoav I HenisPubMed 38168942
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
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Cholesterol/ Cholesteryl Ester Assay Kit ab65359 provides a simple method for sensitive quantification of total cholesterol, free cholesterol, and cholesteryl esters, by colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) plate reader.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
-20°C
- Storage information
-20°C
Notes
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.
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 H2O2 which reacts with a sensitive cholesterol probe to produce 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
Other cholesterol assay kits include: - - C - - Review our to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.
Supplementary info
- Activity summary
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.
- Associated diseases and disorders
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.
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4 product images
Mailloux RJ, Florian M, Chen Q, Yan J, Petrov I, Coughlan MC, et al., PLoS ONE 9(9): e106832., Fig 3, doi.org/10.1371/journal.pone.0106832Functional Studies - Cholesterol/ Cholesteryl Ester Assay Kit - Quantitation (ab65359)
Assessment of free and total cholesterol and cholesteryl ester levels useing ab65359. Protein levels of geranylgeranyldiphosphate synthase were measured by ELISA. Absolute amounts of CoQ10 were measured by HPLC as described in Materials and Methods. Cholesterol levels were quantified using a Cholesterol/Cholesteryl Ester Detection Kit from Abcam according to instruction. n?=?6, means ±SEM. Two-way ANOVA with Tukey's post-hoc test. * denotes statistical comparison between vehicle control (V) and high dose (H) and # denotes statistical comparison between water and ethanol treated groups. * and **, indicate significant difference at p<0.05, 0.01 and 0.0001, respectively. OWV; obese water vehicle, OWH; obese water high dose, OEV; obese ethanol vehicle, OEH; obese ethanol high dose.
Singh, Natesh and Gerhard F Ecker., International journal of molecular sciences vol. 19,5 1278., Fig 1, doi:10.3390/ijms19051278Functional Studies - Cholesterol/ Cholesteryl Ester Assay Kit - Quantitation (ab65359)
Total cholesterol (free and ester forms) were determined with a cholesterol quantification assay (fluorometric detection) (ab65359) and are shown relative to untreated control cells.
Functional Studies - Cholesterol/ Cholesteryl Ester Assay Kit - Quantitation (ab65359)
Cholesterol and cholesteryl ester were quantified colourimetrically in biological fluids (dilution range 1:10-1:100; 50 minutes incubation) and measured in duplicates (+/- SD).
Functional Studies - Cholesterol/ Cholesteryl Ester Assay Kit - Quantitation (ab65359)
Detection of Cholesterol/Cholesteryl Ester Using Cholesterol Quantitation Kit (ab65359). Cholesterol/Cholesteryl Ester was quantified using the kit by colorimetric (A) and fluorometric (B) methods according to the kit instructions. Background from the control reaction (without cholesterol) has been subtracted from each value. Note: Fluorometric assay is over 10 fold more
sensitive than colorimetric assay.
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