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Abcam's Cholesterol Assay Kit (ab65390) provides a simple quantification method of HDL and LDL/VLDL after a convenient separation of HDL from LDL and VLDL (very low-density lipoprotein) in serum samples. In this assay, cholesterol oxidase specifically recognizes free cholesterol and produces a component that will react with probe to generate color (570 nm) and fluorescence (Ex/Em = 538/587 nm). Cholesterol esterase hydrolizes cholesteryl ester into free cholesterol, therefore, cholesterol ester and free cholesterol can be detected separately in the presence and absence of cholesterol esterase in the reactions.
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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.
Regulation of HDL (high-density-lipoprotein)-cholesterol and LDL (low-density-lipoprotein)-cholesterol plays a central role in various disease developments. It is well known that low levels of HDL and high level of LDL are associated with an increased risk of cardiovascular events.
If you need to purchase additional HDL precipitation buffer, please see ab105138.
|2X LDL/VLDL Precipitation Buffer||NW||1 x 10ml|
|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 ug/ul)||Yellow||1 x 100µl|
|Enzyme Mix (Lyophilized)||Green||1 vial|
Our Abpromise guarantee covers the use of ab65390 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent dilution.|
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
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 measure colourimetrically, in duplicates, after 40 minutes.
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).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"