Fluorometric method for the enzymatic determination of cholesterol. 1999

D M Amundson, and M Zhou
Molecular Probes, Inc., Eugene, OR 97402, USA.

A fluorometric method for the enzymatic determination of cholesterol content has been developed using a novel fluorogenic H2O2 probe, Amplex Red. This assay is performed in a 96-well microplate, and it is a one-step method amenable to automated procedures. Using commercially available cholesterol, our assay allows detection of 5 pmol (2 ng) cholesterol per well, which is 100-fold more sensitive than published fluorometric and colorimetric methods. When applied to the measurement of cholesterol levels in serum and food samples, the Amplex Red-based method has been found more attractive since the oxidation product of the Amplex Red method has superior long wavelength spectra which are less susceptible to interference from the biological compounds.

UI MeSH Term Description Entries
D008892 Milk The off-white liquid secreted by the mammary glands of humans and other mammals. It contains proteins, sugar, lipids, vitamins, and minerals. Cow Milk,Cow's Milk,Milk, Cow,Milk, Cow's
D002784 Cholesterol The principal sterol of all higher animals, distributed in body tissues, especially the brain and spinal cord, and in animal fats and oils. Epicholesterol
D002787 Sterol Esterase An enzyme that catalyzes the hydrolysis of CHOLESTEROL ESTERS and some other sterol esters, to liberate cholesterol plus a fatty acid anion. Cholesterol Esterase,15-Ketosteryl Oleate Hydrolase,Acylcholesterol Lipase,Cholesterol Ester Hydrolase,Cholesteryl Oleate Hydrolase,Cholesterylester Hydrolase,Hormone-Sensitive Lipase,Lipase A (Lysosomal Acid Cholesterol Esterase),Lipoidal Steroid Esterase,Lysosomal Acid Cholesterol Esterase,Lysosomal Acid Lipase,Steroid Hormone Esterase,Sterol Ester Acylhydrolase,15 Ketosteryl Oleate Hydrolase,Acid Lipase, Lysosomal,Acylhydrolase, Sterol Ester,Esterase, Cholesterol,Esterase, Lipoidal Steroid,Esterase, Steroid Hormone,Esterase, Sterol,Hormone Sensitive Lipase,Hydrolase, 15-Ketosteryl Oleate,Hydrolase, Cholesterol Ester,Hydrolase, Cholesteryl Oleate,Hydrolase, Cholesterylester,Lipase, Acylcholesterol,Lipase, Hormone-Sensitive,Steroid Esterase, Lipoidal
D004305 Dose-Response Relationship, Drug The relationship between the dose of an administered drug and the response of the organism to the drug. Dose Response Relationship, Drug,Dose-Response Relationships, Drug,Drug Dose-Response Relationship,Drug Dose-Response Relationships,Relationship, Drug Dose-Response,Relationships, Drug Dose-Response
D005456 Fluorescent Dyes Chemicals that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags. Flourescent Agent,Fluorescent Dye,Fluorescent Probe,Fluorescent Probes,Fluorochrome,Fluorochromes,Fluorogenic Substrates,Fluorescence Agents,Fluorescent Agents,Fluorogenic Substrate,Agents, Fluorescence,Agents, Fluorescent,Dyes, Fluorescent,Probes, Fluorescent,Substrates, Fluorogenic
D005470 Fluorometry An analytical method for detecting and measuring FLUORESCENCE in compounds or targets such as cells, proteins, or nucleotides, or targets previously labeled with FLUORESCENCE AGENTS. Fluorimetry,Fluorometric Analysis,Analysis, Fluorometric
D006719 Homovanillic Acid A 3-O-methyl ETHER of (3,4-dihydroxyphenyl)acetic acid. 3-Methoxy-4-Hydroxyphenylacetic Acid,4-Hydroxy-3-Methoxyphenylacetic Acid,3 Methoxy 4 Hydroxyphenylacetic Acid,4 Hydroxy 3 Methoxyphenylacetic Acid,Acid, 3-Methoxy-4-Hydroxyphenylacetic,Acid, 4-Hydroxy-3-Methoxyphenylacetic,Acid, Homovanillic
D000675 Ampyrone A metabolite of AMINOPYRINE with analgesic and anti-inflammatory properties. It is used as a reagent for biochemical reactions producing peroxides or phenols. Ampyrone stimulates LIVER MICROSOMES and is also used to measure extracellular water. Aminoantipyrine,4-Amino-1,5-Dimethyl-2-Phenyl-3H-Pyrazolone,4-Aminoantipyrine,4-Aminophenazone,4 Amino 1,5 Dimethyl 2 Phenyl 3H Pyrazolone,4 Aminoantipyrine,4 Aminophenazone
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity

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