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Glutamine/Glutamate-Glo™ Assay

Sensitive, Quick Detection of Glutamine and Glutamate in Biological Samples

  • Detect even small changes in metabolites
  • Multiplex with other metabolite or viability assays
  • Perform on a variety of sample types

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Catalog number selected: J8021

$ 445.00
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Glutamine/Glutamate-Glo™ Assay
5ml
$ 445.00
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36211300-Energy-Metabolism-image-1280p

Quickly Detect Glutamine and Glutamate from Biological Samples

The Glutamine/Glutamate-Glo™ Assay is a bioluminescent assay for the rapid and sensitive measurement of glutamine and glutamate from a variety of sample types. The bioluminescent signal eliminates signal interference that colorimetric and fluorescent assays suffer. Both glutamine and glutamate are measured from the sample; no separate assay is needed.

How the Assay Works

The assay is based on the conversion of glutamine to glutamate by glutaminase enzyme. Next, glutamate oxidation and NADH production are coupled with a bioluminescent NADH detection system. Glutamate dehydrogenase uses glutamate and NAD+ to produce α-ketoglutarate and 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. 

This assay requires two steps: i) glutamine conversion to glutamate by Glutaminase; and ii) glutamate detection with the Glutamate Detection Reagent. When Glutamate Detection Reagent, which contains glutamate dehydrogenase (GlutDH), NAD+, Reductase, Reductase Substrate and Luciferase, is added to a sample containing glutamate at a 1:1 ratio, the enzyme-coupled reactions start and run simultaneously. The luminescent signal is proportional to the amount of glutamate and increases until all glutamate is consumed, at which time a stable luminescent signal is achieved.

When using this assay, both glutamine and glutamate will be measured. There is no need to use a separate assay for glutamate. For samples that contain both glutamate and glutamine, the light signal will be proportional to the starting concentration of total glutamine plus glutamate. Therefore a second reaction without the Glutaminase enzyme is needed to measure the glutamate-only concentration. Glutamine levels can be calculated by subtracting the glutamate-only signal from the total glutamine plus glutamate signal.

Glutamine and Glutamate Titration Curves

13953MA-W
Glutamine (blue) and glutamate (orange) controls were prepared and diluted as described in Technical Manual TM496. The Glutamine/Glutamate-Glo™ Assay was performed as described and luminescence measured. Because glutaminase efficiently converts glutamine to glutamate, similar RLUs are generated from equal concentrations of glutamine and glutamate.

Advantages

Quickly Measure Metabolites in a Variety of Sample Types: Measure in media, cells, tissue and plasma. The assays require minimal preparation steps with no need for centrifugation and spin columns.

Conduct easy in-well processing for intracellular detection.

Measure Across a Wide Range of Concentrations: Broad linearity up to 3 logs enables simple sample measurement.

Detect Small Changes: Wide assay window with S/Bmax > 300 allows better discrimination of small changes in metabolite levels compared to colorimetric and fluorometric assays.

Gain More Information Per Sample: The assays are amenable to measuring multiple metabolites from the same sample. Multiplex with cell viability assays for normalization.

Glutamine Consumption and Glutamate Secretion by SKOV-3 Cells

13954MA-W-a
13954MA-W-b
As cells grow, they continuously consume glutamine and secrete glutamate. Glutamine consumption (top, blue) and glutamate secretion (bottom, orange) by SKOV-3 ovarian carcinoma cells were monitored over time as described in Technical Manual TM496.

Measure Multiple Metabolites from One Sample

Four metabolites important to the energetic state of the cell—glucose, lactate, glutamate and glutamine—can be measured in parallel using the bioluminescent Glucose-Glo™, Lactate-Glo™, Glutamine/Glutamate-Glo™ and Glutamate-Glo™ Assays. 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.

13836ma-w-a: Glucose consumption measured in metabolite multiplex assay.
13836ma-w-b: Lactate secretion measured in metabolite multiplex assay.
13836ma-w-c: Glutamine consumption measured in metabolite multiplex assay.
13836ma-w-d: Glutamate secretion measured in metabolite multiplex assay.

Specifications

You are viewing: J8021 Change Configuration

What's in the box?

Item Part # SizeConcentration

Reductase

G884A 1 × 55μl6mg/ml

Reductase Substrate

G885A 1 × 55μl

Luciferin Detection Solution

J135A 1 × 5ml

NAD

J136B 1 × 275μl

Glutamate

J139A 1 × 50μl

Glutamate Dehydrogenase

J141A 1 × 100μl

Glutaminase

J142A 1 × 25μl

Glutaminase Buffer

J143A 1 × 2.5ml

SDS

Choose language:

Download SDSPDF (225 KB) – English (United States)

Certificate of Analysis

See all certificates for J8021

Use Restrictions

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

Storage Conditions

AA

Patents and Disclaimers

European Pat. No. 1131441 and Japanese Pat. No. 4520084.

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: J8022 Change Configuration

What's in the box?

Item Part # SizeConcentration

Reductase

G884B 1 × 275μl6mg/ml

Reductase Substrate

G885B 1 × 275μl

Luciferin Detection Solution

J135B 1 × 50ml

NAD

J136C 1 × 1ml

Glutamate

J139A 1 × 50μl

Glutamate Dehydrogenase

J141B 1 × 1ml

Glutaminase

J142B 1 × 125μl

Glutaminase Buffer

J143B 1 × 25ml

SDS

Choose language:

Download SDSPDF (225 KB) – English (United States)

Certificate of Analysis

See all certificates for J8022

Use Restrictions

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

Storage Conditions

AA

Patents and Disclaimers

European Pat. No. 1131441 and Japanese Pat. No. 4520084.

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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