Cholesterol Assay Kit ab65390 is used to quantify total cholesterol, free cholesterol, and cholesterol esters, and includes a method to separate HDL and LDL/VLDL fractions. The assay has one 60 min incubation, with colorimetric (570 nm) or fluorometric (535/587 nm) readout.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Images
Publications
- Nature communications 15:1862024ApoL6 associates with lipid droplets and disrupts Perilipin1-HSL interaction to inhibit lipolysis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYuhui Wang,Hai P Nguyen,Pengya Xue,Ying Xie,Danielle Yi,Frances Lin,Jennie Dinh,Jose A Viscarra,Nnejiuwa U Ibe,Robin E Duncan,Hei S SulPubMed 38167864
- eLife 12:2023Smith-Magenis syndrome protein RAI1 regulates body weight homeostasis through hypothalamic BDNF-producing neurons and neurotrophin downstream signalling.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSehrish Javed,Ya-Ting Chang,Yoobin Cho,Yu-Ju Lee,Hao-Cheng Chang,Minza Haque,Yu Cheng Lin,Wei-Hsiang HuangPubMed 37956053
Key facts
- Detection method
Colorimetric/Fluorometric
- Sample types
Tissue Lysate, Urine, Plasma, Serum, Other biological fluids
- Assay type
Quantitative
- Assay time
1h
Associated Products
Select an associated product type
5 products for Alternative Product
What's included?
Recommended products
Cholesterol Assay Kit ab65390 is used to quantify total cholesterol, free cholesterol, and cholesterol esters, and includes a method to separate HDL and LDL/VLDL fractions. The assay has one 60 min incubation, with colorimetric (570 nm) or fluorometric (535/587 nm) readout.
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
Tissue Lysate, Urine, Plasma, 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 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
Supplementary info
- Activity summary
HDL (High-Density Lipoprotein) and LDL (Low-Density Lipoprotein) often referenced together with VLDL (Very Low-Density Lipoprotein) are essential components in the regulation of cholesterol within the body. HDL known as "good" cholesterol is a spherical particle around 8-11 nm in diameter playing an important role in reverse cholesterol transport. LDL termed "bad" cholesterol and VLDL are involved in delivering cholesterol to cells. These lipoproteins are primarily expressed in the liver and the bloodstream. The LDL length varies from 18-25 nm while VLDL is larger around 30-80 nm. Their importance in cellular cholesterol homeostasis makes them frequent topics of study in lipid metabolism.
- Biological function summary
HDL particles help remove excess cholesterol from peripheral tissues and transport it to the liver for excretion a process important for cardiovascular health. HDL particles are part of a multi-protein complex involving apolipoproteins such as ApoA-I ApoC and ApoE. LDL transports cholesterol to peripheral tissues but when accumulated it can lead to plaque formation in arteries. VLDL primarily acts as a triglyceride carrier converting into LDL after lipid delivery. These lipoproteins function synergistically forming a delicate balance essential for maintaining lipid levels.
- Pathways
HDL and LDL are integral to cholesterol metabolism and energy homeostasis pathways. The reverse cholesterol transport pathway involves HDL in removing cholesterol from macrophages contributing to atheroprotection. LDL is linked to the endocytotic pathway associated with cellular uptake of cholesterol. APOLB is significant in the context of LDL production as it serves as the primary apolipoprotein in VLDL and LDL particles. Interaction between these pathways and lipoproteins determines systemic cholesterol levels and impacts atherosclerotic processes.
- Associated diseases and disorders
Aberrations in HDL and LDL levels are closely associated with cardiovascular diseases like atherosclerosis and hyperlipidemia. An imbalance characterized by high LDL levels or low HDL contributes to plaque buildup in arterial walls leading to heart diseases. HDL and LDL are often modulated in therapeutic strategies aimed at improving cardiovascular health. The PCSK9 protein also plays a vital role where its inhibition leads to increased LDL receptor recycling and decreased blood LDL levels providing a therapeutic target in managing dyslipidemia and related disorders.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
4 product images
Li G et al., PLoS One, 7, e35895, 2012 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Functional Studies - Cholesterol Assay Kit - HDL and LDL/VLDL (ab65390)
Wild type (WT) or farnesoid X receptor knock-out (FRX-KO) mice were treated with either vehicle (Veh) or a high fat-containing diet (HFD) for 16 weeks. Serum HDL (top) and LDL/VLDL cholesterol (bottom) from 5-7 mice/group was measured using ab65390 following protocol instructions. An asterisk (*) means P<0.05 between WT and FXR-KO vehicle group. A pound (#) indicates P<0.05 between Wt and FXR-KO-HFD group. Image obtained from Li G et al., PLoS One, 2012; 7(4): e35895
Functional Studies - Cholesterol Assay Kit - HDL and LDL/VLDL (ab65390)
Measurement of total cholesterol, HDL, LDL/VLDL from serum samples. Total Cholesterol (blue), HDL (green), and LDL/VLDL (cream) cholesterol were measured following the kit protocol.
Functional Studies - Cholesterol Assay Kit - HDL and LDL/VLDL (ab65390)
Rat serum samples were processed according to the protocol. The quantity of total (dilution range 1:10-1:100), HDL (dilution range 1:1-1:10), LDL/VLDL (dilution range 1:1-1:10) and free cholesterol (neat) was measured colourimetrically, in duplicates, after 40 minutes.
Functional Studies - Cholesterol Assay Kit - HDL and LDL/VLDL (ab65390)
Signal from standard curve was measured colourimetrically over a period of time. Background signal subtracted and each point on the curve represents duplicate values (+/- SD).
Downloads
Product protocols
- Visit the general protocols
- Visit troubleshooting
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com