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Pyruvate-Glo™ Assay

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Easy, Sensitive Pyruvate Assay Kit

  • Compatible with cells, media and serum samples
  • Streamlined in-well sample preparation protocol
  • Scalable to 384-well plates for HTS applications

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Catalog number selected: J4051

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Pyruvate-Glo™ Assay
5ml
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Rapid Pyruvate Detection from Diverse Samples

The Pyruvate-Glo™ Assay is a fast and sensitive method for detecting pyruvate in various biological samples, such as cultured cells, media and serum. Based on bioluminescent technology, the Pyruvate-Glo™ Assay can detect subtle changes in glycolysis and mitochondrial metabolism in just 75 minutes. Its streamlined sample preparation protocol involves acid treatment and neutralization directly in the assay wells, eliminating the need for cell collection, centrifugation and spin columns. The assay is adaptable to a 384-well format, facilitating quick analysis of many samples.

How the Pyruvate-Glo™ Assay Works

Diagram illustrating how the Pyruvate-Glo™ Assay works.

Pyruvate Oxidase catalyzes the reduction of pyruvate to acetyl phosphate, generating H2O2. In the presence of H2O2, the H2O2 Substrate is converted into luciferin. This reaction is detected by Ultra-Glo™ Recombinant Luciferase, which emits light proportionate to the amount of pyruvate in the sample.

Simple Assay Protocol Saves Time

Diagram illustrating the Pyruvate-Glo™ Assay protocol workflow.

Accurately Detect the Smallest Changes in Pyruvate

Graph showing the linear relationship between luminescence and pyruvate concentration.



Sensitivity (S/B >5)

1.56μM

Limit of Detection (S/N >3)

400nM

Linear Range

400nM to 50μM

Maximum Assay Window (S/B)

>150

Measure the Effect of Metabolic Inhibitors on Cellular Pyruvate Levels

The Pyruvate-Glo™ Assay can be used to monitor both intracellular and extracellular pyruvate in cell culture, including quickly assessing the metabolic effects of drug treatment.

Bar chart showing the effect on pyruvate levels after adding various drug compounds to K562 cells, as measured by the Pyruvate-Glo™ Assay.
Bar chart showing no decrease in cell viability after incubating K562 cells with various drugs.

Monitor metabolic effects of drug treatment with the  Pyruvate-Glo™ Assay. K562 cells in suspension were incubated with DMSO (negative control), 10μM GSK2837808 (lactate dehydrogenase A inhibitor), 10μM 7ACC2 (monocarboxylate transporter 1-4 [MCT1-4] inhibitor) or 10μM UK5099 (mitochondrial pyruvate carrier inhibitor) for one to two hours. Following the incubations, half the volume was analyzed using the Pyruvate-Glo™ Assay (Panel A), and half the volume was removed for viability analysis using CellTiter-Glo® Luminescent Cell Viability Assay (Panel B). As expected, pyruvate levels increased approximately twofold after a 2-hour incubation with UK5099 and 7ACC2 compared to the DMSO control. There was no decrease in viability during treatment when compared to the DMSO control. All data represent an average of three replicates.

Do you want to learn more about our cellular energy metabolism tools and the innovative technology that powers these assays?

Specifications

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What's in the box?

Item Part # Size

H2O2 Substrate, 10mM

G882A 1 × 40μl

Signal Enhancer Solution

G883A 1 × 100μl

H2O2 Substrate Dilution Buffer

G922A 1 × 2ml

Luciferin Detection Solution

J135A 1 × 5ml

Neutralization Buffer

J153A 1 × 15ml

0.6N HCl

J402A 1 × 15ml

Pyruvate Oxidase (POX)

J416A 1 × 130μl

Pyruvate, 10mM

J417A 1 × 50μl

d-Cysteine, 100X

V251A 1 × 100μl

Certificate of Analysis

Search by lot number

Use Restrictions

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

Storage Conditions

BB

Patents and Disclaimers

Patent Pending.

Specifications

You are viewing: J4052 Change Configuration

What's in the box?

Item Part # Size

H2O2 Substrate, 10mM

G882B 1 × 200μl

Signal Enhancer Solution

G883B 1 × 500μl

H2O2 Substrate Dilution Buffer

G922B 1 × 10ml

Luciferin Detection Solution

J135B 1 × 50ml

Neutralization Buffer

J153A 1 × 15ml

0.6N HCl

J402A 1 × 15ml

Pyruvate Oxidase (POX)

J416B 1 × 1300μl

Pyruvate, 10mM

J417A 1 × 50μl

d-Cysteine, 100X

V251B 1 × 500μl

Certificate of Analysis

Search by lot number

Use Restrictions

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

Storage Conditions

BB

Patents and Disclaimers

Patent Pending.

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