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Lactate-Glo™ Assay

Quick, Sensitive Lactate Assay for a Variety of Biological Samples

  • Perform in high-throughput workflows
  • Multiplex with other metabolite or viability assays
  • Linear lactate detection range up to 200µM

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Catalog number selected: J5021

$ 415.00
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Lactate-Glo™ Assay
5ml
$ 415.00
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Sensitive, Rapid Lactate Detection from a Variety of Samples

The Lactate-Glo™ Assay is a bioluminescent assay for rapid, selective and sensitive detection of L-lactate in biological samples. Lactate is produced by glycolysis, a major metabolic pathway responsible for glucose homeostasis and energy production. Once considered merely a byproduct of glycolysis, lactate is now considered an important regulatory molecule of intermediate metabolism involved in cancer development, diabetes and other diseases.

 

How the Lactate Assay Works

The Lactate-Glo™ Assay couples lactate oxidation and NADH production with a bioluminescent NADH detection system. Lactate dehydrogenase uses lactate and NAD+ to produce pyruvate and NADH. In the presence of NADH a pro-luciferin Reductase Substrate is converted by Reductase to luciferin, which is then used in a luciferase reaction to produce light. The Lactate-Glo™ Assay contains an L-lactate-selective lactate dehydrogenase to confer specificity for L-lactate, the major stereoisomer found in mammalian cells.

When Lactate Detection Reagent, which contains lactate dehydrogenase (LDH), NAD+, Reductase, Reductase Substrate and Luciferase, is added to a sample containing lactate at a 1:1 ratio, the enzyme-coupled reactions start and run simultaneously. The luminescent signal is proportional to the amount of lactate in the sample and increases until all lactate is consumed at which time a stable luminescent signal is achieved.

 

Meets Your Throughput and Workflow Requirements

The Lactate-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. To simplify sample processing, methods for rapid enzyme inactivation and NAD(P)H degradation are provided that are compatible with 96- and 384-well plate formats and do not require sample centrifugation or spin columns.

 

Luminescent Signal is Proportional to Lactate Present in Sample

13887ma-w: Luminescence from lactate assay proportional to metabolite.
Lactate titration curve. Serial dilutions of lactate were prepared as described in TM493, and lactate detected using the Lactate-Glo™ Assay.

Lactate Secretion by A549 Cells

13888ma-w: Luminescent lactate assay measured secretion for 72 hours.
A549 cells were plated at 5,000 (light blue bars) and 15,000 (dark blue bars) cells/well as described in TM493. Red line represents the luminescence of medium-only controls. Samples were taken from the medium at times indicated and the amount of lactate measured using the Lactate-Glo™ Assay. 

Advantages for Lactate-Glo™ Assay

Quickly Detect Lactate in a Variety of Sample Types: Measure in medium, cells, tissue and plasma. The lactose detection assay requires minimal preparation steps with no need for centrifugation and spin columns.

Conduct easy in-well processing for intracellular detection.

Measure Lactate Across a Wide Range of Concentrations: Broad linearity up to 3 logs with simple sample measurement when using the lactate assay.

Detect Small Changes in Lactate Levels: Wide assay window with a S/Bmax > 200 better discriminates small changes in lactate compared to colorimetric and fluorometric lactate assays.

Gain More Information Per Sample: The lactose 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.

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: J5021 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

J136A 1 × 30μl

Lactate

J137A 1 × 50μl

Lactate Dehydrogenase

J195A 1 × 1 each

SDS

Choose language:

Download SDSPDF (174 KB) – English (United States)

Certificate of Analysis

See all certificates for J5021

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: J5022 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

J136B 1 × 275μl

Lactate

J137A 1 × 50μl

Lactate Dehydrogenase

J195A 1 × 1 each

SDS

Choose language:

Download SDSPDF (174 KB) – English (United States)

Certificate of Analysis

See all certificates for J5022

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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