Product nameCholesterol/ Cholesteryl Ester Assay Kit - Quantitation
See all Cholesterol/Cholesteryl Ester kits
Sample typeCell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts
Assay time1h 00m
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 37ºC
- analyze with microplate reader
Other cholesterol assay kits include:
- HDL and LDL/VLDL Cholesterol assay kit ab65390
- Cell-based Cholesterol assay kit ab133116
- Cholesterol Efflux assay kit ab196985
- Cholesterol Uptake assay kit ab236212
Review our Metabolism Assay Guide 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.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 100 tests Cholesterol Assay Buffer WM 1 x 25ml Cholesterol Esterase (lyophilized) Blue 1 vial Cholesterol Probe (in DMSO, anhydrous) Red 1 x 200µl Cholesterol Standard (2 µg/µl ) Yellow 1 x 100µl Enzyme Mix (lyophilized) Green 1 vial
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Cholesterol Metabolism
RelevanceCholesterol is an essential molecule in all animal life. It is involved in both normal development and diseases. A high level of serum cholesterol is an indicator for diseases such as heart disease. Cholesteryl ester is an ester of cholesterol. The ester bond is formed between the carboxylate group of a fatty acid and the hydroxyl group of cholesterol.
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.
Total cholesterol (free and ester forms) were determined with a cholesterol quantification assay (fluorometric detection) (ab65359) and are shown relative to untreated control cells.
Cholesterol and cholesteryl ester were quantified colourimetrically in biological fluids (dilution range 1:10-1:100; 50 minutes incubation) and measured in duplicates (+/- SD).
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.
This product has been referenced in:
- Tong XK et al. High cholesterol triggers white matter alterations and cognitive deficits in a mouse model of cerebrovascular disease: benefits of simvastatin. Cell Death Dis 10:89 (2019). Read more (PubMed: 30692517) »
- Jo Y et al. Schnyder corneal dystrophy-associated UBIAD1 inhibits ER-associated degradation of HMG CoA reductase in mice. Elife 8:N/A (2019). Read more (PubMed: 30785396) »