Absolute quantitation of intracellular metabolite concentrations by an isotope ratio-based approach

@article{Bennett2008AbsoluteQO,
  title={Absolute quantitation of intracellular metabolite concentrations by an isotope ratio-based approach},
  author={Bryson D. Bennett and Jie Yuan and Elizabeth H Kimball and Joshua D. Rabinowitz},
  journal={Nature Protocols},
  year={2008},
  volume={3},
  pages={1299-1311}
}
This protocol provides a method for quantitating the intracellular concentrations of endogenous metabolites in cultured cells. The cells are grown in stable isotope-labeled media to near-complete isotopic enrichment and then extracted in organic solvent containing unlabeled internal standards in known concentrations. The ratio of endogenous metabolite to internal standard in the extract is determined using mass spectrometry (MS). The product of this ratio and the unlabeled standard amount… CONTINUE READING
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