title: One-step procedure for peptide extraction from in-gel digestion sample for mass spectrometric analysis.
abstract: Protein identification from samples resolved by one-dimensional and two-dimensional gel electrophoresis is highly dependent on the recovery of trypsin-digested peptides prior to mass spectrometric analysis. The commonly used two-step protocol for extracting tryptic peptides, involving high-volume organic solvent extraction and cleanup via microscale reversed-phase micropipet tip or microcolumn is not only limited by significant sample loss but is also costly and very labor-intensive. We report here a simple one-step procedure for simultaneous peptide extraction and cleanup by incubating a small piece of C18 Empore Disk (3M) with the in-gel digested solution. We show that the direct Empore Disk-based peptide extraction procedure is convenient, economical, and has higher efficiency as compared with the commonly used two-step protocol for peptide preparation prior to MS analysis.
authors.affiliation: School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551.
authors.forename: Wei
authors.initials: W
authors.lastname: Meng
authors.forename: Huoming
authors.initials: H
authors.lastname: Zhang
authors.forename: Tiannan
authors.initials: T
authors.lastname: Guo
authors.forename: Chhiti
authors.initials: C
authors.lastname: Pandey
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhu
authors.forename: Oi Lian
authors.initials: OL
authors.lastname: Kon
authors.forename: Siu Kwan
authors.initials: SK
authors.lastname: Sze
mesh_terms: D015180:Electrophoresis, Gel, Two-Dimensional; D029968:Escherichia coli Proteins / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry*; D020543:Proteome / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D010446:Peptide Fragments; D020543:Proteome; D014357:Trypsin