Large-scale phosphosite quantification in tissues by a spike-in SILAC method

@article{Monetti2011LargescalePQ,
  title={Large-scale phosphosite quantification in tissues by a spike-in SILAC method},
  author={Mara Monetti and Nagarjuna Nagaraj and Kirti Sharma and Matthias Mann},
  journal={Nature Methods},
  year={2011},
  volume={8},
  pages={655-658}
}
Despite progress in mass spectrometry (MS)-based phosphoproteomics, large-scale in vivo analyses remain challenging. Here we report a 'spike-in' stable-isotope labeling with amino acids in cell culture (SILAC) methodology using standards derived from labeled mouse liver cell lines, using which we analyzed insulin signaling. With this approach we identified 15,000 phosphosites and quantitatively compared 10,000 sites in response to insulin treatment, creating a very large, accurately quantified… CONTINUE READING

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