 The Cholesterol/Cholesteryl Ester Quantitation Kit provides a simple method for sensitive quantification of cholesterol, cholesteryl Ester, or both by either colorimetric or fluorometric methods. A large portion of the cholesterol in blood is in the form of cholesteryl esters. Cholesterol esterase hydrolizes cholesteryl ester into cholesterol. Cholesterol is then oxidized by cholesterol oxidase to yield H2O2. The produced H2O2 interacts with a sensitive cholesterol probe to produce resorufin, which can be detected by spectrophotometry at  = 570 nm or fluorometry at Ex/Em = 535/587 nm. The assay can detect cholesterol itself (without adding cholesterol esterase) or total cholesterol (cholesterol + cholesteryl ester) by adding cholesterol esterase to the reaction, or cholesteryl Ester itself by subtracting the value of cholesterol from the total value of cholesterol and cholesteryl esters.
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Detection of Cholesterol/Cholesteryl Ester Using Cholesterol Quantitation Kit. Cholesterol/Cholesteryl Ester was quantified using the Cholesterol probe by colorimetric (A) and fluorometric (B) methods according to the kit instructions. Background from the control reaction (without cholesterol added) has been subtracted from each value.
Note: Fluorometric assay is 4-10 folds more sensitive than colorimetric assay.
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The regulation of HDL (high-density-lipoprotein)-cholesterol and LDL (low-density-lipoprotein)-cholesterol plays a central role in various disease development. It is well known that low levels of HDL and high level of LDL are associated with an increased risk of cardiovascular events. The HDL and LDL/VLDL Cholesterol Quantification Kit provides a simple method for convenient separation of HDL from LDL and VLDL (very low-density lipoprotein) in serum sample, then HDL and LDL/VLDL cholesterol levels can easily be quantified separately, using cholesterol oxidase mediated reactions. Cholesterol esterase hydrolizes cholesteryl ester into free cholesterol, which can then be oxidized by cholesterol oxidase to yield H2O2. The H2O2 interacts with a sensitive cholesterol probe to produce resorufin, which can be detected with a micro-plate reader at l = 570 nm or fluorescence micro-plate reader at Ex/Em = 538/587 nm. The assay can also detect cholesterol ester and free cholesterol separately in these two fractions by adding or not adding cholesterol esterase into the reactions.
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Measurement of total cholesterol, HDL, LDL/VLDL from serum samples. Total Cholesterol (dark bar), HDL (shadow Bar), and LDL/VLDL (open Bar) cholesterol was measured follow the kit protocol.
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