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.

In the cholesterol assay protocol, cholesterol oxidase acts on free cholesterol to produce a chemical which reacts with a probe to generate color (570 nm) and fluorescence (Ex/Em = 538/587 nm). Cholesterol esterase is used to hydrolyze cholesteryl ester into free cholesterol.

If cholesterol esterase is included in the assay, total cholesterol is measured. If it is not included, free cholesterol is measured. The amount of cholesterol ester can be calculated by subtracting free cholesterol from total cholesterol.

Note: 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.

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

Other cholesterol assay kits include: - - - - 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. **How other researchers have used Cholesterol Assay Kit ab65390** This 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* serum*7* 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