Increased proteome coverage by combining PAGE and peptide isoelectric focusing: Comparative study of gel-based separation approaches

@inproceedings{Atanassov2013IncreasedPC,
  title={Increased proteome coverage by combining PAGE and peptide isoelectric focusing: Comparative study of gel-based separation approaches},
  author={Ilian Atanassov and Henning Urlaub},
  booktitle={Proteomics},
  year={2013}
}
The in-depth analysis of complex proteome samples requires fractionation of the sample into subsamples prior to LC-MS/MS in shotgun proteomics experiments. We have established a 3D workflow for shotgun proteomics that relies on protein separation by 1D PAGE, gel fractionation, trypsin digestion, and peptide separation by in-gel IEF, prior to RP-HPLC-MS/MS. Our results show that applying peptide IEF can significantly increase the number of proteins identified from PAGE subfractionation. This… CONTINUE READING