Cayman’s Cholesterol Fluorometric/Colorimetric Assay Kit provides a simple method for the sensitive quantitation of cholesterol in biological samples, such as plasma, serum, cell extracts, and tissue extracts. The assay is based on an enzyme-coupled reaction that detects both free cholesterol and cholesteryl esters. Cholesteryl esters are hydrolyzed by cholesterol esterase into cholesterol, which is then oxidized by cholesterol oxidase to yield hydrogen peroxide and the corresponding ketone product. By excluding cholesterol esterase, only free cholesterol will be oxidized and therefore can be distinguished from total cholesterol. Hydrogen peroxide produced by cholesterol oxidase activity is then detected using MaxiProbe, a highly sensitive and stable probe for hydrogen peroxide.1 In the presence of horseradish peroxidase, MaxiProbe reacts with hydrogen peroxide with a 1:1 stoichiometry to produce a compound, the fluorescence of which can be quantified at excitation and emission wavelengths of 530 and 590 nm, respectively. Alternatively, its absorbance can be measured at 570 nm. The fluorometric assay has a range of 0-20 μM with a lower limit of detection of 1 μM. The colorimetric assay has a range of 0-100 μM with a lower limit of detection of 1 μM.Needed but not supplied: Please download the kit booklet to verify if UltraPure Water (Milli-Q or equivalent) or any other components are needed for this assay.WARNING This product is not for human or veterinary use.
