Cholesterol/Cholesterol Ester-Glo™ Assay

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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

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Catalog number selected: J3190

$ 595.00
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Cholesterol/Cholesterol Ester-Glo™ Assay
5ml
$ 595.00
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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.

Diagram showing how cholesterol and cholesterol ester are detected with the cholesterol-glo assay.

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 mouse liver using a cholesterol spike.

Cholesterol Detection in Human Lipoproteins

Cholesterol detection in human lipoproteins using the Cholesterol-glo assay.
Purchased human high density and low density lipoproteins were assayed for both total (samples + esterase) and free cholesterol. Concentrations were calculated using a 40μM cholesterol standard. Relative light unit (RLU) values (in millions) are listed at the top of each bar.

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.

Specifications

You are viewing: J3190 Change Configuration

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

Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

AA

Patents and Disclaimers

U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.

Specifications

You are viewing: J3191 Change Configuration

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

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

AA

Patents and Disclaimers

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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Alessandro

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