title: IPG strip-based peptide fractionation for shotgun proteomics.
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.
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin, Thielallee 63, 14195, Berlin, Germany.
authors.forename: Murat
authors.initials: M
authors.lastname: Eravci
authors.forename: Christian
authors.initials: C
authors.lastname: Sommer
authors.forename: Matthias
authors.initials: M
authors.lastname: Selbach
mesh_terms: D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D057809:HEK293 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D010455:Peptides / Q000302:isolation & purification*; D040901:Proteomics
chemical_list: D010455:Peptides