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Extraction and Solubilization of Proteins for Proteomic Studies

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The Proteomics Protocols Handbook

Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

For any proteomic study involving various control and experimental specimens, several factors need to be in place. A critical one is the extraction and solubilization of all components, regardless of whether a chromatographic (1,2) or two-dimensional (2-D) gel electrophoretic fractionation (3–6) is performed prior to analysis of proteins of interest by mass spectrometry of protein digests. All proteins must not only be extracted, but they must also be completely soluble, free from interacting partners (such as protein-RNA/DNA and protein-protein interactions, metabolites, and so on), and, in the case of 2-D gel electrophoresis, they must remain soluble as they approach their isoelectric points. The solubilization process should extract all classes of proteins reproducibly, such that statistically significant quantitative data can be obtained and correlated with experimental perturbations and the resulting biological responses.

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  1. PGRD-World Wide Safety Sciences Pfizer, Inc., St.Louis, MO

    Richard M. Leimgruber

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  1. University of Hertfordshire, Hatfield, UK

    John M. Walker

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Leimgruber, R.M. (2005). Extraction and Solubilization of Proteins for Proteomic Studies. In: Walker, J.M. (eds) The Proteomics Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1385/1-59259-890-0:001

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  • DOI: https://doi.org/10.1385/1-59259-890-0:001

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-343-5

  • Online ISBN: 978-1-59259-890-8

  • eBook Packages: Springer Protocols

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