GeLC-MS/MS analysis of complex protein mixtures

Methods Mol Biol. 2014;1156:53-66. doi: 10.1007/978-1-4939-0685-7_4.

Abstract

Discovery-based proteomics has found its place in nearly every facet of biological research. A key objective of this approach is to maximize sequence coverage for proteins across a wide concentration range. Fractionating samples at the protein level is one of the most common ways to circumvent challenges due to sample complexity and improve proteome coverage. Of the available methods, one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by liquid chromatography-tandem mass spectrometry (GeLC-MS/MS) is a robust and reproducible method for qualitative and quantitative proteomic analysis. Here we describe a general GeLC-MS/MS protocol and include technical advice and outline caveats to increase the probability of a successful analysis.

MeSH terms

  • Chromatography, Liquid / methods*
  • Complex Mixtures
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Proteins / chemistry*
  • Reproducibility of Results
  • Tandem Mass Spectrometry / methods*

Substances

  • Complex Mixtures
  • Proteins