Pyruvate, which is produced in the final step of glycolysis, participates in anabolic metabolism, catabolic metabolism, and signal transduction. Invitrogen Molecular Probes Amplex UltraRed Reagent 5 x 1mg Electrophoresis, Western Blotting and ELISA:Western Blotting Reagents, Membranes. Invitrogen Molecular Probes Amplex Red/UltraRed Stop Reagent 1 set Life Sciences:Cell Analysis:Cell Based Assays:Cell Growth and Differentiation Reagents.

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The results show that Amplex UltraRed improves the performance of pyruvate assay, providing enhanced signal sensitivity. Each point is the mean of triplicate measurements.

Amplex UltraRed enhances the sensitivity of fluorimetric pyruvate detection

Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

To overcome this problem, we examined this fluorimetric assay with the new dye Amplex UltraRed Cat. Error bars are not shown because their sizes are smaller than the symbols. Free Radic Biol Med. Pyruvate, which is produced maplex the final step of glycolysis, participates in anabolic metabolism, catabolic metabolism, and signal transduction.

Analysis of High-throughput screening data from Amplex UltraRed (AUR) Thyroperoxidase (TPO) assays

The NaClO 4 concentration is A simple statistical parameter for use in evaluation and validation of high throughput screening assays. An excitation wavelength of nm is used.

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Please review our privacy policy. We’ve made some changes to EPA. Academic Press; New York: The final reaction solution contained mM potassium phosphate with 1. Therefore, the enzymatic reactions are favorable in acidic buffer, while the fluorogenic reaction is more efficient in a neutral or weakly basic solution.

Regulation of store-operated calcium channels by the intermediary metabolite pyruvic eed. Our results show that Amplex UltraRed improves the performance of the assay, providing brighter fluorescence and enhanced sensitivity. The role of phosphometabolites in cell proliferation, energy metabolism, and tumor therapy.

Concentrations of metabolites uotra animal tissues. However, its background fluorescence is higher. Abstract Pyruvate, which is produced in the final step of glycolysis, participates in anabolic metabolism, catabolic metabolism, and signal transduction. Due to a lapse in appropriations, EPA websites will not be regularly updated.

Pyruvate protects neurons against hydrogen peroxide-induced toxicity. Author manuscript; available in PMC Aug 1. Please note that all information on the EPA website may not amplfx up to date, and transactions and inquiries submitted to the EPA website may not be processed or responded to until appropriations are enacted. For bacterial pyruvate oxidase, one unit will produce 1. An official website of the United States government.

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The background intensity was subtracted using the value of the no pyruvate control for all sample readings. The pH optimum of HRP is in the range of 6. Pyruvate is produced by pyruvate kinase EC2. In addition, H 2 O 2 has been reported to cause a nonenzymatic and stoichiometric decarboxylation of pyruvate [ 56 ].

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In this case, the measured intensity is reduced by changing the excitation wavelength to nm where the linearity of the standard curve is improved in comparison to nm Fig.

However, the absorption and fluorescence of resorufin are pH-dependent. See other articles in PMC that cite the published article. Support Center Support Center. The side reaction between H 2 O 2 and pyruvate can be effectively suppressed in acidic buffer pH 5.

As pyruvate production is associated with cell growth control, energy production, and intracellular signaling [ 12 ], it is important to have sensitive assays to evaluate pyruvate concentrations.

The first column in Fig. As HClO 4 is commonly used as a deproteinizing reagent in pyruvate sample preparation, we tested its effect on this assay.

Group members share an interest in encouraging computational toxicology usage and exposure science in helping rred implement EPA’s mission of protecting human health and the environment. A sensitive fluorimetric assay for pyruvate.

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