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Optimization of the procedure of nitrogen oxide quantitation in mammalian cell culture media

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Abstract

A procedure for quantitative analysis of nitric oxide production by cultured mammalian cells based on nitrite ion quantitation has been optimized. The Griess assay adapted for microvolumes was found to yield optimal results when the NO −3 ions formed from NO −2 ions in the reaction medium were reduced by nonactivated cadmium prior to the assay to form the analyte NO −2 . The assay was verified for the culture of C6 rat glioma cells. The assay developed is suitable for quantitative determination of nitrogen oxide in 96- and 48-well cell culture plates; the detection limit for NO −2 was 2.1 ± 0.1 μM and that for NO −3 was 2.9 ± 0.1 μM.

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Abbreviations

NED:

N 1-naphthylethylendiamine dihydrochloride

SA:

sulfanilamide

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Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya, 16/10, Moscow, 117997, Russia

    M. G. Akimov, E. V. Fomina-Ageeva & V. V. Bezuglov

Authors
  1. M. G. Akimov
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  2. E. V. Fomina-Ageeva
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  3. V. V. Bezuglov
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Correspondence to M. G. Akimov.

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Original Russian Text © M.G. Akimov, E.V. Fomina-Ageeva, V.V. Bezuglov, 2015, published in Bioorganicheskaya Khimiya, 2015, Vol. 41, No. 1, pp. 74–81.

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Akimov, M.G., Fomina-Ageeva, E.V. & Bezuglov, V.V. Optimization of the procedure of nitrogen oxide quantitation in mammalian cell culture media. Russ J Bioorg Chem 41, 63–69 (2015). https://doi.org/10.1134/S1068162015010021

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  • DOI: https://doi.org/10.1134/S1068162015010021

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