Glycogen-Glo™ Assay

Sensitive In-Well Detection of Glycogen

The Glycogen-Glo™ Assay is a rapid, sensitive method for detecting glycogen in biological samples. The assay allows you to prepare samples and read signal from the same well without additional cell collection, centrifugation or spin columns. Based on bioluminescent technology, the Glycogen-Glo™ Assay can detect subtle changes in glycogen synthesis, storage and breakdown.

How the Glycogen-Glo™ Assay Works

How to detect glycogen using the Glycogen-Glo assay. — The Glycogen-Glo™ Assay utilizes a series of enzyme-coupled reactions. First, glycogen is digested into glucose using glucoamylase enzyme. The resulting glucose is then measured using glucose dehydrogenase in conjunction with a bioluminescent NADH detection technology. The result is a light signal proportional to the starting glycogen concentration in the sample.

How to Determine Glycogen Concentration

Samples that contain both glycogen and glucose require two reactions to determine the glycogen concentration. One reaction is used to measure total glucose resulting from digested glycogen plus any glucose present in the sample. The second is used to measure any glucose that was present in the sample prior to digestion. The difference in signal between the two reactions represents the contribution from glycogen in the sample. The effective digestion of glycogen into glucose monomers (≥70%) simplifies data interpretation and calculations.

Simple Detection Protocol

Detect the Smallest Changes in Glycogen

Bioluminescent detection provides superior sensitivity compared to colorimetric and fluorescent glycogen assays, so fewer cells are needed to determine intracellular glycogen levels.

Sensitivity	<1µg/ml
Limit of Detection	<20ng/ml
Linear Range	To 20µg/ml glycogen
Maximum Assay Window (S/B)	>250

Data showing the sensitive detection of glycogen using the Glycogen-Glo assay. — Bioluminescent signal (RLU) is proportional to glycogen content of the sample and dependent on glucoamylase enzyme treatment.

Monitor Glycogen Depletion and Accumulation in Cells

Monitoring glycogen depletion and accumulation using the Glycogen-Glo Assay. — Preparation of cell lysates for the Glycogen-Glo™ assay is simple and can be performed in-well. To prepare lysates for the following experiments, the medium was removed and the cells were washed three times with PBS before lysing with acid (HCl). **Panel A. Cells deplete glycogen during starvation.** A total of 12,500 cells were grown overnight in culture medium containing 25mM glucose or starvation medium containing 0mM glucose. **Panel B. Starved cells accumulate glycogen in the presence of glucose.** Cells were starved overnight, collected and then plated in medium containing 0 or 10mM glucose for 24 hours.

Kit Sizes

5ml kit (sufficient for 100 assays in 96-well plate)
50ml kit (sufficient for 1,000 assays in 96-well plate)

Reaction volumes can be scaled easily to 384-well plates for high-throughput applications.

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

SDS

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Certificate of Analysis

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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.

SDS

Search for SDS

Certificate of Analysis

Search by lot number

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

Bioluminescent Assays for Metabolism

Other Resources

Frequently Used With

GloMax® Discover System

High-performance microplate reader for detecting luminescence, fluorescence and absorbance.

GM3000

GloMax® Explorer System

Easy-to-use, flexible multimode detection system.

GM3500, GM3510

Glycogen-Glo™ Assay

Easy, Sensitive Glycogen Assay Kit for Biological Samples

Sensitive In-Well Detection of Glycogen

How the Glycogen-Glo™ Assay Works

How to Determine Glycogen Concentration

Simple Detection Protocol

Detect the Smallest Changes in Glycogen