title: GeLC-MS/MS analysis of complex protein mixtures.
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.
authors.affiliation: Biochemistry and Molecular Genetics, University of Colorado Denver, Aurora, CO, 80045, USA.
authors.forename: Monika
authors.initials: M
authors.lastname: Dzieciatkowska
authors.forename: Ryan
authors.initials: R
authors.lastname: Hill
authors.forename: Kirk C
authors.initials: KC
authors.lastname: Hansen
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D045424:Complex Mixtures; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D045424:Complex Mixtures; D011506:Proteins