title: In-gel peptide IEF sample preparation for LC/MS analysis.
abstract: The technique of proteolytically digesting a sample and identifying its protein components by liquid chromatography followed by mass spectrometry (LC-MS) is a widely used analytical tool. Prior fractionation by isoelectric focusing (IEF) may be performed to increase the depth of proteome coverage. Here, we describe a method for in-gel IEF separation of a proteolytic digest that utilizes commercially available immobilized pH gradient (IPG) strips and a widely used IEF instrument.
authors.affiliation: Bio-Rad Laboratories, Life Science Group, Hercules, CA, USA, Thomas_berkelman@bio-rad.com.
authors.forename: Tom
authors.initials: T
authors.lastname: Berkelman
authors.forename: Sricharan
authors.initials: S
authors.lastname: Bandhakavi
authors.forename: Aran
authors.initials: A
authors.lastname: Paulus
mesh_terms: D002853:Chromatography, Liquid; D006367:HeLa Cells; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification; D052616:Solid Phase Extraction
chemical_list: D010455:Peptides