EnzyChrom™ Pyruvate Assay Kit
ProtocolSDS


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Application
For quantitative determination of pyruvate and evaluation of drug effects on its metabolism.

Key Features
Sensitive and accurate. Use as little as 10 μL samples. Linear detection range in 96-well plate: 2 to 500 μM (17 μg/dL to 4.4 mg/dL) pyruvate for colorimetric assays and 0.2 to 50 μM for fluorimetric assays.

Simple and convenient. The procedure involves addition of a single working reagent and incubation for 30 min at room temperature, compatible for HTS assays.

Improved reagent stability. The optimized formulation has greatly enhanced the reagent and signal stability.

Method
OD570nm, or FL530/585nm

Samples
Biological

Species
All

Procedure
20 min

Size
100 tests

Detection Limit
OD, FL: 2, 0.2 μM

Shelf Life
6 months

More Details
Pyruvate is a key intermediate in cellular metabolic pathways. Pyruvate can be converted to carbohydrates via gluconeogenesis, to fatty acids or energy through acetyl-CoA, to the amino acid alanine and to ethanol. Abnormal levels of pyruvate have been linked to liver diseases and metabolic disorders. Simple, direct and automation-ready procedures for measuring pyruvate concentrations find wide applications in research and drug discovery. BioAssay Systems pyruvate assay uses a single Working Reagent that combines pyruvate oxidase and hydrogen peroxide determination in one step. The color intensity of the reaction product at 570nm or fluorescence intensity at λex/em = 530/585nm is directly proportional to pyruvate concentration in the sample.

1. I would like to use the EnzyChrom pyruvate assay kit (EYPR-100) to determine pyruvate levels in cell culture medium containing phenol red and fetal bovine serum. Does this interfere in any way with the measurement?



Phenol red adds a background to the OD measurement that will vary with the pH of the medium. Pyruvate in FBS will also add a background to the measurement. In order to correct for those interferences you should add a separate control with unused medium +FBS and subtract that from all sample values or, you could also prepare the standard in the medium + FBS. To play it safe. Alternatively you can use serum-free medium without phenol red.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
1. Verma MK et al (2014). Integrated analysis of chronic lipotoxicity on muscle metabolism and stress and its reversal by antioxidants. Springerplus. 3: 251. Assay: Pyruvate in Human cell lysate (Pubmed).

2. Wu J et al (2014). Pyruvate-associated acid resistance in bacteria. Appl Environ Microbiol. 80(14):4108-13. Assay: Pyruvate in E. Coli cells (Pubmed).

3. Cui Z, Gao N, Wang Q, Ren Y, Wang K, Zhu T (2015). BcMctA, a putative monocarboxylate transporter, is required for pathogenicity in Botrytis cinerea. Curr Genet. [Epub ahead of print] Assay: Pyruvate in Hyphae Tissue (Pubmed).

4. Darekar S, et al (2012). Epstein-Barr virus immortalization of human B-cells leads to stabilization of hypoxia-induced factor 1 alpha, congruent with the Warburg effect. PLoS One. 7(7): e42072. Assay: Pyruvate in Human cells (Pubmed).

5. Park MR, et al (2012). Transcriptome profiling characterizes phosphate deficiency effects on carbohydrate metabolism in rice leaves. J Plant Physiol. 169(2):193-205. Assay: Pyruvate in Plant leaves (Pubmed).

6. Cajuday, LA (2014). Carbohydrate Uptake in Water Buffalo Cumulus-Oocyte Complexes (COCs) Supplemented with Retinoic Acid During In Vitro Maturation. Philippine Journal of Science. 143(1): 43-48.

7. Fong, JE et al (2013). Moderate excess of pyruvate augments osteoclastogenesis. Biol Open. 2(4):387-95. Assay: Serum in Mice (Pubmed).

8. Chen, X et al (2015). Terazosin activated Pgk1 and Hsp90 to promote stress resistance. Nat Chem Biol. 11(1):19-25. Assay: Cells in Flies (Pubmed).

9. Yasid, NAB (2013). Metabolite analysis of Escherichia coli in response to changes in oxygen levels. Department of Molecular Biology and Biotechnology, The University of Sheffield.

To find more recent publications, please click here.
Please inquire or request assay service or call 1-510-782-9988 x 2.
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EnzyChrom™ Pyruvate Assay Kit
Catalog No: EPYR-100
Price: $319    Qty:
For orders of 10 or more kits, please call 1-510-7829988x1 or email us for best pricing and/or bulk order.

Shipping: On Ice
Shipment: Fedex Service
Delivery: 1-2 days (US), 3-6 days (Intl) Storage: -20°C

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