Description
How the Assay Works
The Glucose-Glo™ Assay couples glucose oxidation and NADH production with a bioluminescent NADH detection system. Glucose dehydrogenase uses glucose and NAD+ to produce NADH. In the presence of NADH a pro-luciferin Reductase Substrate is converted by Reductase to luciferin that is then used by Ultra-Glo™ Recombinant Luciferase to produce light.
When Glucose Detection Reagent, which contains glucose dehydrogenase (GDH), NAD+, Reductase, Reductase Substrate and Luciferase, is added to a sample containing glucose at a 1:1 ratio, the enzyme-coupled reactions start and run simultaneously. The luminescent signal is proportional to the amount of glucose in the sample and increases until all glucose is consumed, at which time a stable luminescent signal is achieved.
Luminescent Signal is Proportional to Amount of Glucose Present
Amenable to Your Throughput and Workflow Requirements
The Glucose-Glo™ Assay is a versatile detection kit that is amenable to high-throughput formats and compatible with many sample types including cells, lysates, tissue, plasma and serum. To simplify sample processing, the glucose detection kit uses processing methods that do not require sample centrifugation or spin columns. Simply dilute your samples, add inactivation and neutralization solutions (if needed), and assay all samples directly in 96- or 384-well plates.
Glucose Consumption by Adherent Lung Carcinoma A549 Cells
Measure Glycolysis and Glutaminolysis from One Sample
Easily measure changes in cellular glycolysis and glutaminolysis by sampling medium over time. Direct measurement of four metabolites important to the energetic state of the cell—glucose, lactate, glutamate and glutamine—can be performed in parallel using the bioluminescent Glucose-Glo™, Lactate-Glo™, Glutamine/Glutamate-Glo™ and Glutamate-Glo™ Assays. No specialized instrument, plates or artificial medium is required. Sample processing compatible with all of the bioluminescent metabolite assays means the same sample can be used for detecting all four metabolites. This includes sample types such as culture medium, serum, plasma and tissue. The data below were produced by measuring metabolites using medium samples from the same cell cultures as described in Technical Manual TM494.
Advantages for Glucose-Glo™ Assay
Quickly Detect Glucose in a Variety of Sample Types: Measure in medium, cells, tissue and plasma. The glucose detection assay requires minimal preparation steps with no need for centrifugation and spin columns.
Detect Small Changes in Glucose Levels Across a Wide Range of Concentrations : Sensitive glucose assay with broad linearity better discriminates small changes in glucose compared to colorimetric and fluorometric glucose assays.
Gain More Information Per Sample: The glucose detection assay along with the other bioluminescent metabolite detection assays are amenable to measuring multiple metabolites from the same sample. Multiplex with cell viability assays for normalization.
Requires Only a Luminometer: The same plate reader that measures cell viability can be used to monitor glycolysis. No specialized instruments, plates or artificial medium are required.
Further Reading About Glucose-Glo™ Assay Development
Zhou, W. et al. (2014) Self-immolative bioluminogenic quinone luciferins for NAD(P)H assays and reducing capacity-based cell viability assay s. Chem BioChem 15 , 670–5.
Vidugiriene, J. et al . (2014) Bioluminescent cell-based NAD(P)/NAD(P)H assays for rapid dinucleotide measurement and inhibitor screening . Assay and Drug Development Technologies 12 , 514–26.
Leippe, D. et al . (2017) Bioluminescent assays for glucose and glutamine metabolism: High-throughput screening for changes in extracellular and intracellular metabolites . SLAS Discovery 22 (4), 366–77.
