One-step procedure for peptide extraction from in-gel digestion sample for mass spectrometric analysis

Wei Meng, Huoming Zhang, Tiannan Guo, Chhiti Pandey, Yi Zhu, Lian Kon Oi, Kwan Sze Siu

    Research output: Contribution to journalArticlepeer-review

    14 Scopus citations

    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.

    Original languageEnglish (US)
    Pages (from-to)9797-9805
    Number of pages9
    JournalAnalytical Chemistry
    Volume80
    Issue number24
    DOIs
    StatePublished - Dec 15 2008

    ASJC Scopus subject areas

    • Analytical Chemistry

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