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Cholesterol Assay Kit (ab65390) provides a simple quantification method for HDL and LDL/VLDL cholesterol after a convenient separation of HDL from LDL and VLDL in mammalian serum samples.
In this cholesterol assay, cholesterol oxidase specifically 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 also used to hydrolyze 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 assay.
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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|
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).
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