The Amplex® Red Cholesterol Assay Kit provides a sensitive, rapid, and simple fluorometric method for detecting very low concentrations of cholesterol using a fluorescence microplate reader or fluorometer. The assay measures both free cholesterol and cholesteryl esters.
• Detects as little as 200 nM (80 ng/ml) cholesterol• Helps accurately measure cholesterol content of 0.01 µl of human serum• Detects both free cholesterol and cholesteryl esters• Allows for multiple time point measurements• Designed for minimal autofluorescence interference
The Amplex Red Cholesterol Assay kit contains these reagents and buffers:
• Amplex Red reagent
• Horseradish peroxidase
• Hydrogen peroxide
• Cholesterol esterase
• Cholesterol oxidase
• Cholesterol reference standard
• Resorufin fluorescence reference standard
• 5X Reaction bufferCholesterol Measurement is Simple and Fast
Performing the Amplex® Red cholesterol assay is quick and easy, requiring only a 30-minute incubation after addition of reagents with no separation steps required. The 100 µl assay volume is designed for 96-well plates; fluorescence can be measured with a microplate reader or fluorometer. Since the assay is continuous, once the Amplex® Red reagent is added, multiple time-point measurements can be taken. The kit is complete with all necessary reagents except deionized water.Helps Accurately Detect Very Low Cholesterol Concentrations
The Amplex® Red cholesterol assay utilizes enzyme-coupled reactions to produce highly fluorescent resorufin for quantitation. Resorufin is produced by the reaction of the Amplex® Red reagent with H2
produced from the cholesterol oxidase-catalyzed oxidation of cholesterol. Because a large proportion of cholesterol in blood is in the form of cholesteryl esters, cholesterol esterase is used to produce free cholesterol from cholesterol esters. Performing the assay in the presence and absence of cholesterol esterase potentially allows determination of the fractions of cholesterol in each form.
This highly sensitive method allows the Amplex® Red Cholesterol Assay kit to detect concentrations of cholesterol as low as 200 nM and measure the cholesterol content of 0.01 µl of human serum. Because the absorbance and emission maxima of resorufin are 571 nm and 585 nm respectively, autofluorescence in most biological samples does not interfere with the assay.Use Amplex® Red Assays for a Broad Range of Investigations
A wide variety of validated Invitrogen Amplex® Red assays are available for studying cell signaling and lipids, neurobiology, inflammation and immune function, and metabolism. We also offer Amplex® UltraRed (Cat. No. A36006), a second-generation reagent providing greater sensitivity and brighter fluorescence and the Amplex® Red/UltraRed stop reagent (Cat. No. A33855). The Amplex® Red/UltraRed stop reagent provides convenience and control by allowing the fluorescence signal-generating reaction to be terminated at a user-determined time point. After addition of the stop reagent, the fluorescence signal remains stable for at least 3 hours. Custom assay design and packaging are also available.
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.Related Link
• Learn More about Amplex® Red Assays and Technology