Pyruvate-Glo™ Assay
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
Catalog Number:
Size
Catalog Number: J4051
Catalog Number: J4052
Rapid Pyruvate Detection from Diverse Samples
Pyruvate-Glo™ アッセイは、培養細胞、培地、血清など様々な生体試料中のピルビン酸を迅速かつ高感度に検出する方法です。生物発光技術を採用した Pyruvate-Glo™ アッセイは、解糖とミトコンドリア代謝により起こるわずかな変化を約75分で検出できます。その合理化されたサンプル調製プロトコールは、酸処理と中和をアッセイウエル内で直接行うため、細胞採取、遠心分離、スピンカラムが不要です。このアッセイは384ウェルフォーマットに適応可能で、多くのサンプルの迅速な分析を可能にします。
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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
Accurately Detect the Smallest Changes in Pyruvate
|
Sensitivity (S/B >5) |
1.56μM |
|
|
Limit of Detection (S/N >3) |
400nM |
|
|
Linear Range |
400nM to 50μM |
|
|
Maximum Assay Window (S/B) |
>150 |
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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.
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.
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Protocols
Complete Protocol
Specifications
Catalog Number:
選択製品の構成品内容
| 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 |
選択製品の構成品内容
| 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 |
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