title: Capillary isoelectric focusing--mass spectrometry of proteins and protein complexes.
abstract: Complex proteome samples require efficient separation and detection methods in order to characterize their protein components. On-line combination of capillary isoelectric focusing (CIEF) with electrospray ionization (ESI) mass spectrometry (MS) is shown as an effective method to analyze complex protein mixtures. Our experience with several microorganisms allowed us to establish successful experimental protocol. Here we use the example of E. coli whole-cell lysate for the CIEF separation and MS detection on the intact protein level. The protocol was further adapted for the analysis of the mixture of noncovalent complexes on the intact complex level.
authors.affiliation: Environmental and Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA., USA.
authors.forename: Suzana
authors.initials: S
authors.lastname: Martinović
authors.forename: Ljiljana
authors.initials: L
authors.lastname: Pasa-Tolić
authors.forename: Richard D
authors.initials: RD
authors.lastname: Smith
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000032:analysis*; D007525:Isoelectric Focusing / Q000379:methods; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D029968:Escherichia coli Proteins