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Proteomics in Neural Crest Cell Research

  • Peter McCarthy
  • Quenten SchwarzEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1976)

Abstract

Detailed investigation of the neural crest cell proteome has lingered behind that of the transcriptome due to the challenge of obtaining sufficient starting material for subsequent proteomic investigation. Compounded by the complexity of protein abundance, large number of posttranslational modifications, and the stochastic nature of proteomics approaches, little data so far exists describing the true neural crest cell proteome. However, recent advances in instrument sensitivity and recovery of material during sample preparation have alleviated many of these problems and make proteomics analysis an underutilized tool to study neural crest cell biology. Here we present a quantitative proteomics protocol for deep analysis of both whole proteome and posttranslational modifications and a separate protocol for ultrasensitive proteomic profiling from submicrogram amounts of protein.

Key words

Neural crest Proteomics SILAC Proteome 

References

  1. 1.
    Vogel C, Marcotte EM (2012) Insights into the regulation of protein abundance from proteomic and transcriptomic analyses. Nat Rev Genet 13(4):227–232.  https://doi.org/10.1038/nrg3185CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Calderon-Celis F, Encinar JR, Sanz-Medel A (2017) Standardization approaches in absolute quantitative proteomics with mass spectrometry. Mass Spectrom Rev.  https://doi.org/10.1002/mas.21542CrossRefGoogle Scholar
  3. 3.
    Geiger T, Cox J, Ostasiewicz P, Wisniewski JR, Mann M (2010) Super-SILAC mix for quantitative proteomics of human tumor tissue. Nat Methods 7(5):383–385.  https://doi.org/10.1038/nmeth.1446CrossRefPubMedGoogle Scholar
  4. 4.
    Hughes CS, Foehr S, Garfield DA, Furlong EE, Steinmetz LM, Krijgsveld J (2014) Ultrasensitive proteome analysis using paramagnetic bead technology. Mol Syst Biol 10:757.  https://doi.org/10.15252/msb.20145625CrossRefPubMedGoogle Scholar
  5. 5.
    Wilson YM, Richards KL, Ford-Perriss ML, Panthier JJ, Murphy M (2004) Neural crest cell lineage segregation in the mouse neural tube. Development 131(24):6153–6162.  https://doi.org/10.1242/dev.01533CrossRefPubMedGoogle Scholar
  6. 6.
    Wiszniak S, Kabbara S, Lumb R, Scherer M, Secker G, Harvey N, Kumar S, Schwarz Q (2013) The ubiquitin ligase Nedd4 regulates craniofacial development by promoting cranial neural crest cell survival and stem-cell like properties. Dev Biol 383(2):186–200.  https://doi.org/10.1016/j.ydbio.2013.09.024CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Centre for Cancer BiologyUniversity of South Australia and SA PathologyAdelaideAustralia

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