Cholesterol/Cholesterol Ester-Glo™ Assay
Detect Cholesterol from a Variety of Biological Samples
- Works with lysed cells, tissues, medium and serum samples
- No organic extraction, extreme heat or centrifugation required
- Linear cholesterol detection up to 80µM
Catalog Number:
Size
Catalog Number: J3190
Catalog Number: J3191
Sensitive, Rapid Detection of Cholesterol and Cholesterol Esters from a Variety of Samples
The Cholesterol/Cholesterol Ester-Glo™ Assay is a bioluminescent assay for rapid and sensitive method for measuring cholesterol and cholesterol esters in cultured cell lysates and other biological samples, such as lipoprotein fractions, cell culture medium, serum and tissue homogenates. Cholesterol is an essential lipid involved in steroidogenesis, bile acid synthesis, cell signaling and maintenance of membrane structure.
How the Assay Works
The Cholesterol/Cholesterol Ester-Glo™ Assay measures cholesterol using a cholesterol dehydrogenase that links the presence of cholesterol to the production of NADH and the activation of a proluciferin that produces light with luciferase. In the presence of NADH, Reductase enzymatically reduces a proluciferin Reductase Substrate to luciferin. Luciferin is detected in a luciferase reaction using Ultra-Glo™ Luciferase and ATP, and the amount of light produced is proportional to the amount of cholesterol in the sample.
For measuring Cholesterol Ester, the assay includes a cholesterol esterase that will remove the fatty acid from cholesterol esters to produce one molecule of cholesterol per molecule of ester. The amount of cholesterol esters is determined from the difference of cholesterol measured in the absence (free cholesterol) and presence (total cholesterol) of esterase.
Simple Protocol is Compatible with Diverse Sample Types
Cholesterol Detection in Mouse Liver Tissue
Mouse liver tissue was homogenized and diluted in Lysis Solution. A cholesterol standard spike of 10μM was added to an aliquot of diluted sample, then all samples and standard were assayed for free cholesterol. The change between standard and background is the same as the change between sample + standard and sample, indicating that the tissue homogenate did not interfere with quantitation of the cholesterol standard.
Cholesterol Detection in Human Lipoproteins
Fractions of lipoproteins contain different amounts of cholesterol and cholesterol esters. The cholesterol detection assay can be used to quantify total cholesterol levels, free cholesterol levels and calculate the cholesterol ester levels as the difference between the two values.
Protocols
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size | Concentration |
|---|---|---|---|
Reductase Substrate |
G885A | 1 × 55μl | |
Cholesterol Lysis Solution |
J319A | 1 × 10ml | |
Cholesterol Detection Solution |
J320A | 1 × 5ml | |
Cholesterol Esterase |
J321A | 1 × 50μl | |
Cholesterol Standard |
J322A | 1 × 500μl | 20mM |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
What's in the box?
| Item | Part # | Size | Concentration |
|---|---|---|---|
Reductase Substrate |
G885B | 2 × 275μl | |
Cholesterol Lysis Solution |
J319B | 1 × 100ml | |
Cholesterol Detection Solution |
J320B | 1 × 50ml | |
Cholesterol Esterase |
J321B | 1 × 500μl | |
Cholesterol Standard |
J322A | 1 × 500μl | 20mM |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
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