One-step procedure for peptide extraction from in-gel digestion sample for mass spectrometric analysis.
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.
Meng
Wei
W
School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551.
Zhang
Huoming
H
Guo
Tiannan
T
Pandey
Chhiti
C
Zhu
Yi
Y
Kon
Oi Lian
OL
Sze
Siu Kwan
SK
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
D029968:Escherichia coli Proteins; D010446:Peptide Fragments; D020543:Proteome; D014357:Trypsin