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Glutamate-Glo™ Assay

Detect Glutamate from a Variety of Biological Samples

  • Amenable to high-throughput formats
  • Detect even intracellular levels of glutamate
  • Limit of detection in the nM range

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Catalog number selected: J7021

$ 415.00
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Glutamate-Glo™ Assay
5ml
$ 415.00
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Bioluminescent Detection of Glutamate from Biological Samples

Glutamate is an important metabolite, serving as a precursor for the synthesis of nucleic acids, nucleotides and proteins. Glutamate is also a key neurotransmitter in nerve cells. Upregulated glutamate production is used as a marker of increased cancer cell dependence on glutaminolysis to support high proliferation rates. The Glutamate-Glo™ Assay is a suitable assay for measuring changes in glutamate levels in a variety of samples, and the assay is sensitive enough to detect even intracellular amounts of glutamate.

How the Assay Works

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. 

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 in the sample and increases until all glutamate is consumed, at which time a stable luminescent signal is achieved.

Luminescent Signal is Proportional to Glutamate Present

13899MA-W
Glutamate titration curve. Dilutions of glutamate were prepared as described in Technical Manual TM495, and the Glutamate-Glo™ Assay was performed.

Flexible Throughput and Process

The Glutamate-Glo™ Assay is a versatile system that is amenable to high-throughput formats and compatible with many sample types. Samples may require upfront sample processing, including dilutions, to fit into the linear range of the assay. They may also require inactivation of endogenous enzyme activity/deproteinization and NAD(P)H degradation (recommendations in technical manual). To simplify sample processing, methods for rapid enzyme inactivation and NAD(P)H degradation are provided that are compatible with 96- and 384-well plates and do not require sample centrifugation or spin columns.


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 > 100 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.

Glutamate Secretion by SKOV-3 Cells

13900MA-W
SKOV-3 ovarian carcinoma cells were plated at 5,000 (light blue bars) or 15,000 (dark blue bars) cells/well. Medium was sampled at indicated time points, and Glutmate-Glo™ Assay was performed as described in Technical Manual TM495.

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: J7021 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

SDS

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Download SDSPDF (176 KB) – English (United States)

Certificate of Analysis

See all certificates for J7021

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: J7022 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

SDS

Choose language:

Download SDSPDF (176 KB) – English (United States)

Certificate of Analysis

See all certificates for J7022

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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