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Proteomics for Biomarker Discovery pp 295–309Cite as

Label-Free Differential Analysis of Murine Postsynaptic Densities

Part of the Methods in Molecular Biology book series (MIMB,volume 1002)

Abstract

This chapter provides detailed methodology for the enrichment and label-free differential analysis of postsynaptic density (PSD) proteins. Methods discussed will include tissue homogenization, subcellular fractionation, protein digestion, and label-free differential analysis after liquid chromatography–tandem mass spectrometry. When combined, these protocols facilitate the identification of receptors and signal transducers that comprise the PSD and provide an optimized workflow for the differential analysis of PSD proteomes. This strategy supports a utility for coupling fractionation with proteomics analysis to enrich for low-abundant proteins in cellular localizations that would otherwise be lost in a global tissue context.

Key words

  • Postsynaptic density
  • Label-free
  • Differential analysis
  • Neuroproteomics
  • Tandem mass spectrometry
  • Sucrose gradient centrifugation
  • Topograph

The online version of the original chapter can be found at http://dx.doi.org/10.1007/978-1-62703-360-2_24

An erratum to this chapter can be found at http://dx.doi.org/10.1007/978-1-62703-360-2_24

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Acknowledgments

This work was supported by the following grants: HD41697-10 (SPG), GM103533/AA016171(MJM), and AA016653/AA016171 (CCW).

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Authors and Affiliations

  1. University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    Scott P. Goulding & Christine C. Wu

  2. University of Washington, Seattle, WA, USA

    Michael J. MacCoss

Authors
  1. Scott P. Goulding
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  2. Michael J. MacCoss
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  3. Christine C. Wu
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Editor information

Editors and Affiliations

  1. , Laboratory of Proteomics and Analytical, Frederick National Laboratory for Cancer, N/A, Frederick, 21702-1201, Maryland, USA

    Ming Zhou

  2. , Laboratory of Proteomics and Analytical, Frederick National Laboratory for Cancer, N/A 5540, Frederick, 21702-1201, Maryland, USA

    Timothy Veenstra

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Goulding, S.P., MacCoss, M.J., Wu, C.C. (2013). Label-Free Differential Analysis of Murine Postsynaptic Densities. In: Zhou, M., Veenstra, T. (eds) Proteomics for Biomarker Discovery. Methods in Molecular Biology, vol 1002. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-360-2_22

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  • DOI: https://doi.org/10.1007/978-1-62703-360-2_22

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-359-6

  • Online ISBN: 978-1-62703-360-2

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