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IPG strip-based peptide fractionation for shotgun proteomics

Authors

  • M. Eravci
  • C. Sommer
  • M. Selbach

Journal

  • Methods in Molecular Biology

Citation

  • Methods Mol Biol 1156: 67-77

Abstract

  • Efficient fractionation of peptides is an essential prerequisite for comprehensive analysis of complex protein mixtures by shotgun mass spectrometry. The separation of peptides by isoelectric focusing is particularly attractive due to its orthogonality to reverse-phase HPLC. Here, we present a protocol for in-gel peptide isoelectric focusing using immobilized pH gradient strips. The method shows high resolving power for up to 1 mg of sample and is highly reproducible.


DOI

doi:10.1007/978-1-4939-0685-7_5