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MaxQuant for In-Depth Analysis of Large SILAC Datasets

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1188))

Abstract

Proteomics experiments can generate very large volumes of data, in particular in situations where within one experimental design many samples are compared to each other, possibly in combination with pre-fractionation of samples prior to LC-MS analysis. Here we provide a step-by-step protocol explaining how the current MaxQuant version can be used to analyze large SILAC-labeling datasets in an efficient way.

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References

  1. Cox J, Mann M (2007) Is proteomics the new genomics? Cell 130:395–398

    Article  CAS  PubMed  Google Scholar 

  2. Cox J, Mann M (2011) Quantitative, high-resolution proteomics for data-driven systems biology. Annu Rev Biochem 80:273–299

    Article  CAS  PubMed  Google Scholar 

  3. de Godoy LM, Olsen JV, Cox J et al (2008) Comprehensive mass-spectrometry-based proteome quantification of haploid versus diploid yeast. Nature 455:1251–1254

    Article  PubMed  Google Scholar 

  4. Ong SE, Mann M (2006) A practical recipe for stable isotope labeling by amino acids in cell culture (SILAC). Nat Protoc 1:2650–2660

    Article  CAS  PubMed  Google Scholar 

  5. Ong SE, Blagoev B, Kratchmarova I et al (2002) Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics. Mol Cell Proteomics 1:376–386

    Article  CAS  PubMed  Google Scholar 

  6. Mann M (2006) Functional and quantitative proteomics using SILAC. Nat Rev Mol Cell Biol 7:952–958

    Article  CAS  PubMed  Google Scholar 

  7. Geiger T, Wisniewski JR, Cox J et al (2011) Use of stable isotope labeling by amino acids in cell culture as a spike-in standard in quantitative proteomics. Nat Protoc 6:147–157

    Article  CAS  PubMed  Google Scholar 

  8. Geiger T, Cox J, Ostasiewicz P et al (2010) Super-SILAC mix for quantitative proteomics of human tumor tissue. Nat Methods 7:383–385

    Article  CAS  PubMed  Google Scholar 

  9. Cox J, Mann M (2008) MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat Biotechnol 26:1367–1372

    Article  CAS  PubMed  Google Scholar 

  10. Cox J, Mann M (2009) Computational principles of determining and improving mass precision and accuracy for proteome measurements in an Orbitrap. J Am Soc Mass Spectrom 20:1477–1485

    Article  CAS  PubMed  Google Scholar 

  11. Cox J, Neuhauser N, Michalski A et al (2011) Andromeda: a peptide search engine integrated into the MaxQuant environment. J Proteome Res 10:1794–1805

    Article  CAS  PubMed  Google Scholar 

  12. Cox J, Mann M (2012) 1D and 2D annotation enrichment: a statistical method integrating quantitative proteomics with complementary high-throughput data. BMC Bioinform 13(Suppl 16):S12

    Article  CAS  Google Scholar 

  13. Luber CA, Cox J, Lauterbach H et al (2010) Quantitative proteomics reveals subset-specific viral recognition in dendritic cells. Immunity 32:279–289

    Article  CAS  PubMed  Google Scholar 

  14. Cox J, Michalski A, Mann M (2011) Software lock mass by two-dimensional minimization of peptide mass errors. J Am Soc Mass Spectrom 22:1373–1380

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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Correspondence to Jürgen Cox .

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Tyanova, S., Mann, M., Cox, J. (2014). MaxQuant for In-Depth Analysis of Large SILAC Datasets. In: Warscheid, B. (eds) Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). Methods in Molecular Biology, vol 1188. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1142-4_24

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  • DOI: https://doi.org/10.1007/978-1-4939-1142-4_24

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1141-7

  • Online ISBN: 978-1-4939-1142-4

  • eBook Packages: Springer Protocols

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