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 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
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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 products
Cholesterol Assay Kit - HDL and LDL/VLDL ab65390 is identical to Cholesterol/
Cholesteryl Ester Assay Kit Cholesterol/
Cholesteryl Ester Assay Kit - Quantitationab65359, except for the addition
of a buffer and protocol for separation of HDL and LDL/VLDL fractions.
Getting the best performance from ab65390
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.
How other researchers are using 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
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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 products
Other cholesterol assay kits include:
Cholesterol assay kit ab65390 is often used with
Triglyceride assay kit Triglyceride
Assay Kit - Quantificationab65336, and Free Fatty Acid assay kit Free
Fatty Acid Assay Kit - Quantificationab65341, to study lipid metabolism and
its role in various metabolic diseases such as diabetes, obesity, liver
diseases, and cardiovascular conditions.
Learn more about our tools for obesity research, including antibodies and ELISA
kits to adipogenesis and lipid metabolism markers, tools for GLP-1 receptor
agonist research, and enzymatic assays to key biochemicals and enzymes involved
in obesity-related metabolism.
Other notes
This 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.
Our broad range of gold-standard reagents features highly
specific recombinant antibodies and kits, validated across applications and
species to ensure the precision and reproducibility your work requires. Drive
your drug discovery efforts forward with our tailored
solutions designed to meet the unique challenges of obesity research.
Abcam has not and does not intend to apply for the REACH
Authorisation of customers' uses of products that contain European
Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application
of REACH Authorisation, and any other relevant authorisations, for their
intended uses.
