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- W2001077777 abstract "Stable isotope, and in particular 13C-based flux analysis, is the exclusive approach to experimentally quantify the integrated responses of metabolic networks. Here we describe a protocol that is based on growing microbes on 13C-labeled glucose and subsequent gas chromatography mass spectrometric detection of 13C-patterns in protein-bound amino acids. Relying on publicly available software packages, we then describe two complementary mathematical approaches to estimate either local ratios of converging fluxes or absolute fluxes through different pathways. As amino acids in cell protein are abundant and stable, this protocol requires a minimum of equipment and analytical expertise. Most other flux methods are variants of the principles presented here. A true alternative is the analytically more demanding dynamic flux analysis that relies on 13C-pattern in free intracellular metabolites. The presented protocols take 5–10 d, have been used extensively in the past decade and are exemplified here for the central metabolism of Escherichia coli." @default.
- W2001077777 created "2016-06-24" @default.
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- W2001077777 date "2009-05-21" @default.
- W2001077777 modified "2023-10-11" @default.
- W2001077777 title "13C-based metabolic flux analysis" @default.
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- W2001077777 doi "https://doi.org/10.1038/nprot.2009.58" @default.
- W2001077777 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/19478804" @default.
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