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Extraction and Solubilization of Proteins for Proteomic Studies

  • Richard M. Leimgruber
Protocol
Part of the Springer Protocols Handbooks book series (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, 4, 5, 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.

Keywords

Sodium Dodecyl Sulfate Bromphenol Blue Linear Ramp Carrier Ampholyte Zwitterionic Detergent 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc., Totowa, NJ 2005

Authors and Affiliations

  • Richard M. Leimgruber
    • 1
  1. 1.PGRD-World Wide Safety Sciences Pfizer, Inc.

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