Cholesterol Assay Kit - HDL and LDL/VLDL

Cholesterol Assay Kit ab65390 uses a simple method to quantify total cholesterol, free cholesterol, and cholesterol esters in mammalian samples. It also includes an easy method to separate HDL and LDL / VLDL cholesterol. How the assay worksThe 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 - Use complete sample for cholesterol measurement; or to separate HDL and LDL/VLDL, mix sample with precipitation buffer, incubate for 10 min, centrifuge for 10 min at 2,000 g, keep HDL fraction supernatant, repeat centrifugation and resuspend LDL/VLDL fraction pellet.- For assay, 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 productsCholesterol 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. Getting the best performance from ab65390Note: This kit is primarily designed for use with serum and plasma samples. It has been successfully used with liver samples (including in Chang et al. 2019, PMID: 30670059, Yu et al. 2018, PMID: 29867579 and Sadh et al. 2017, PMID: 28800633); sample preparation in liver (and other tissue) samples may need optimization and modification by the user for successful use. How other researchers are using Cholesterol assay kit ab65390This cholesterol assay kit has been used in publications in a variety of sample types, including: - Human: serum1- Mouse: plasma and liver tissue2, plasma3, serum4,1- Rat: liver tissue5, serum1, plasma6- Fish: Z. temminckii serum7 References: 1 - Lin C et al 2018; 2 - Change RC et al 2019; 3 - Hinder et al 2019, Havighorst A et al 2019, Guo X et al 2017, Wen et al 2018, Fernandes GW et al 2018, Ahnstedt H et al 2018, Dumas SN and Ntambi JM 2018; 4 - Liu J et al 2018, Cui XB et al 2019, Rohm et al 2018; 5 - Yu S et al 2018; 6 - Park JB 2017; 7 - Chae Y et al 2018 Related and recommended productsOther cholesterol assay kits include: - Cell-based Cholesterol assay kit ab133116- Cholesterol Efflux assay kit ab196985- Cholesterol Uptake assay kit ab236212 Review our metabolism assays overview to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress here.  Other notesThis product is manufactured by BioVision, an Abcam company and was previously called K613 HDL and LDL/VLDL Quantification Colorimetric/Fluorometric Kit. K613-100 is the same size as the 100 test size of ab65390.

其他

Quantitative

Serum, Plasma, Other biological fluids, Tissue Lysate

High density lipoprotein LDL Low density lipoprotein Very low density lipoprotein VLDL see all

Microplate reader