Glycerol-Glo™ Assay

Rapid and Sensitive Glycerol Detection in Biological Samples

The Glycerol-Glo™ Assay is a bioluminescent assay for rapid and sensitive measurement of glycerol in a variety of biological samples, including cells grown in monolayer or 3D structures, cell culture medium, tissues and serum samples. Glycerol is often measured as the product of lipolysis, where it is released from triglycerides. Glycerol is also a substrate or product of many other enzymatic or metabolic processes that can be studied with the glycerol assay. Any processes that result in changes in glycerol concentration, both extracellular and intracellular, can be studied with the Glycerol-Glo™ Assay.

How the Assay Works

The Glycerol-Glo™ Assay measures glycerol in a coupled reaction scheme that links the production of NADH to the activation of a proluciferin that produces light with luciferase. Glycerol kinase and glycerol-3-phosphate dehydrogenase are used to generate NADH.

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 glycerol in the sample.

Diagram showing how the glycerol-glo assay detects glycerol

Simple, Flexible Sample Preparation

Glycerol Release from Adipocytes

The Glycerol-Glo™ Assay can be used with cultured cells, including 3D cell culture models, homogenized tissue samples, cell culture medium samples and serum or plasma samples.

No organic extraction is required. The Glycerol Lysis Reagent effectively extracts glycerol from your sample in a single reagent addition without extreme heat, centrifugation steps or multi-step organic extraction protocols.

Glycerol release from adipocytes detected using the Glycerol-glo assay. — Adipocytes were differentiated from 3T3L1-MBX fibroblasts. Medium was removed and the cells were washed twice with PBS. Medium containing 2% fatty acid-free BSA and 5μM triacsin C to promote extracellular glucose accumulation was added, with or without 10μM isoproterenol to induce stimulated or control conditions, respectively. At the indicated time points, aliquots of medium were removed and assayed for glycerol. The glycerol detection assay is used to quantitate lipolysis in adipocytes.

Protocols

Complete Protocol

Glycerol-Glo™ Assay Technical ManualPDF (2062 KB) – English

Quick Protocols

Glycerol-Glo Assay FB228PDF (246 KB) – English

What's in the box?

Item	Part #	Size	Concentration
Reductase Substrate	G885A	1 × 55μl
Glycerol Lysis Solution	J315A	1 × 10ml
Kinetic Enhancer	J316A	1 × 50μl
Glycerol Detection Solution	J317A	1 × 5ml
Glycerol Standard	J324A	1 × 500μl	20mM

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

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

Storage Conditions

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.

What's in the box?

Item	Part #	Size	Concentration
Reductase Substrate	G885B	2 × 275μl
Glycerol Lysis Solution	J315B	1 × 100ml
Kinetic Enhancer	J316B	1 × 500μl
Glycerol Detection Solution	J317B	1 × 50ml
Glycerol Standard	J324A	1 × 500μl	20mM

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

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

Storage Conditions

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.

Resources

Articles

Homogeneous Assays for Triglyceride Metabolism Research

Posters

Other Resources

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Simple, Sensitive Glycerol Detection Assay for a Variety of Biological Samples

Rapid and Sensitive Glycerol Detection in Biological Samples

How the Assay Works