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Quenching Methods for the Analysis of Intracellular Metabolites

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Animal Cell Biotechnology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1104))

Abstract

Sampling and quenching methods for intracellular metabolite analysis in mammalian cells in adherent and suspension culture are described. Quenching of adherent cells is achieved by application of hot air after removal of the supernatant by suction. For suspension cultures, the addition of excess ice-cold saline results in a rapid inactivation of metabolism and significant dilution of extracellular metabolites. Medium carryover is prevented by rinsing the cells with washing solution. Separation of supernatant from suspension cells via centrifugation is incomplete due to required short centrifugation time. Thus, it is necessary to determine the reproducible cell recovery after quenching.

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Author information

Authors and Affiliations

  1. Biochemical Engineering, Saarland University, Saarbrücken, Germany

    Judith Wahrheit & Elmar Heinzle

Authors
  1. Judith Wahrheit
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  2. Elmar Heinzle
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Editors and Affiliations

  1. Bioprozess- und Biosystemtechnik, Technische Universität Hamburg-Harburg, Hamburg, Germany

    Ralf Pörtner

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Wahrheit, J., Heinzle, E. (2014). Quenching Methods for the Analysis of Intracellular Metabolites. In: Pörtner, R. (eds) Animal Cell Biotechnology. Methods in Molecular Biology, vol 1104. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-733-4_14

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  • DOI: https://doi.org/10.1007/978-1-62703-733-4_14

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-732-7

  • Online ISBN: 978-1-62703-733-4

  • eBook Packages: Springer Protocols

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