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In Vivo Quantitative Proteomics: The SILAC Mouse

  • Sara Zanivan
  • Marcus Krueger
  • Matthias Mann
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 757)

Abstract

Mass spectrometry-based proteomics is a field that has been quickly developing, enabling increasingly giving in-depth characterization of the proteomes of cells and tissues. Current technology allows identifying thousands of proteins in a single experiment. Stable isotope labeling with amino acid in cell culture (SILAC) was originally developed for high accuracy quantitative proteomic studies in cell lines. We have shown that SILAC can be extended to in vivo animal model by fully labeling C57BL/6 mice with 13C6-Lysine (Lys6). We used SILAC mice technology to map quantitative proteomic changes in mice lacking the expression of β1 integrin, β-Parvin, or the integrin tail-binding protein Kindlin-3. This approach confirmed the absence of the proteins and revealed a role of Kindlin-3 in red blood cells. Here we describe a practical method to generate and maintain a colony of SILAC mice and optimal strategies to perform in vivo quantitative proteomic experiments.

Key words

SILAC mouse Mass spectrometry In vivo quantitative proteomics Integrins Cell adhesion 

Notes

Acknowledgments

The authors would like to thank SILANTES for the development of Lys6-containing mouse diet and Marcus Moser for contributing to the application of the SILAC mouse technology.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Vascular Proteomics GroupBeatson Institute for Cancer ResearchGlasgowUK
  2. 2.Department of Cardiac Development and RemodelingMax-Planck-Institute of Heart and Lung ResearchBad NauheimGermany
  3. 3.Department of Proteomics and Signal TransductionMax-Planck-Institute of Heart and Lung ResearchBad NauheimGermany

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