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Invitrogen™ Amplex™ Red Cholesterol Assay Kit, 500 Assays
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Specifications:
| Application | Fluorescence-Based Assay | ||
| Storage Temperature | -5 to -30°C | ||
| Product Type | Assay Kits | Forms | Kit with Various components |
| Product Brand | Thermo Fisher Scientific™, Invitrogen | ||
| Product Grade | Molecular Biology | ||
The Invitrogen™ Amplex™ Red Cholesterol Assay Kit provides a highly sensitive, rapid, and easy-to-use fluorometric method for detecting very low concentrations of cholesterol in biological samples. Compatible with both microplate readers and fluorometers, the assay can measure free cholesterol, cholesteryl esters, or both.
This kit delivers detection sensitivity down to 200 nM (80 ng/mL) and enables measurement of cholesterol content in as little as 0.01 μL of human serum, making it ideal for lipid metabolism studies, cardiovascular research, and biochemical screening.
Assay Principle
The assay uses an enzyme-coupled reaction to produce the red-fluorescent oxidation product resorufin:
- Cholesterol esterase hydrolyzes cholesteryl esters to free cholesterol.
- Cholesterol oxidase oxidizes cholesterol to cholestenone, producing hydrogen peroxide (H₂O₂).
- In the presence of horseradish peroxidase (HRP), H₂O₂ reacts with the Amplex™ Red reagent to generate resorufin.
Resorufin’s excitation/emission maxima (571/585 nm) minimize autofluorescence interference from most biological samples.
Key Features
- Ultra-Sensitive Detection – Detects as little as 200 nM cholesterol.
- Measures Both Cholesterol Forms – Quantifies free cholesterol and cholesteryl esters.
- Fast, Simple Workflow – 30-minute incubation; no separation steps required.
- Continuous Assay Capability – Allows multiple time-point measurements.
- Low Background Interference – Minimal autofluorescence impact.
- Complete Kit – All reagents included except deionized water.
Applications
- Serum and Plasma Cholesterol Measurement
- Lipid Metabolism Research
- Cardiovascular and Atherosclerosis Studies
- Drug Screening for Cholesterol Modulators
- Metabolic Disorder Investigations
- Basic Biochemistry and Enzymology Studies
Technical Specifications
| Parameter | Specification |
|---|---|
| Detection Method | Fluorescence |
| Sensitivity | 200 nM (80 ng/mL) cholesterol |
| Analytes | Free cholesterol, cholesteryl esters |
| Format | 96-well plate |
| Excitation/Emission | 571 / 585 nm |
| Sample Volume | As low as 0.01 μL serum |
| Assay Time | ~30 minutes |
| Kit Size | 500 assays |
| Storage | –5°C to –30°C, protect from light |
| Shipping | Room temperature |
Kit Contents
- Amplex™ Red reagent
- DMSO
- Horseradish peroxidase (HRP)
- Hydrogen peroxide
- Cholesterol esterase
- Cholesterol oxidase
- Cholesterol reference standard
- Resorufin fluorescence reference standard
- 5X reaction buffer
The Invitrogen™ Amplex™ Red Cholesterol Assay Kit is an ideal solution for precise, sensitive, and reproducible cholesterol quantification. Its ability to measure both free and esterified cholesterol with minimal sample volume makes it a valuable tool in cardiovascular, metabolic, and biochemical research.