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2-D Proteome Analysis Protocols pp 571–588Cite as

Peptide Sequencing of 2-DE Gel-Isolated Proteins by Nanoelectrospray Tandem Mass Spectrometry

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 112))

Abstract

Shortly after the introduction of electrospray as a viable ionization technique for large molecules (1), electrospray tandem mass spectrometry (ES MS/MS) techniques, such as HPLC-ES MS/MS, were used successfully for peptide sequencing at picomole and subpicomole levels (2–4). In LC-MS/MS, peptide sequence information is generated during the short time, 10–30 s, that a peptide elutes from the HPLC column run at a flow rate of 0.5–5 μL/min. This time frame rarely allows optimization of experimental parameters for MS/MS sequencing of individual peptides unless several LC-MS/MS experiments can be performed on a sample. With the introduction of the nanoelectrospray ion source (5–7), the time constraint of tandem mass spectrometry has been removed, and peptide sequencing has been reliably extended to the femtomole level of gel-isolated protein.

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Author information

Authors and Affiliations

  1. Protein Research Group, Department of Molecular Biology, Odense University, Odense, Denmark

    Ole N. Jensen

  2. Protein and Peptide Group, European Molecular Biology Laboratory, Heidelberg, Germany

    Matthias Wilm, Andrej Shevchenko & Matthias Mann

Authors
  1. Ole N. Jensen
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  2. Matthias Wilm
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  3. Andrej Shevchenko
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  4. Matthias Mann
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Editor information

Editors and Affiliations

  1. Department of Molecular Biotechnology, University of Washington, Seattle, WA

    Andrew J. Link

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© 1999 Humana Press Inc., Totowa, NJ

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Jensen, O.N., Wilm, M., Shevchenko, A., Mann, M. (1999). Peptide Sequencing of 2-DE Gel-Isolated Proteins by Nanoelectrospray Tandem Mass Spectrometry. In: Link, A.J. (eds) 2-D Proteome Analysis Protocols. Methods in Molecular Biology, vol 112. Humana Press. https://doi.org/10.1385/1-59259-584-7:571

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  • DOI: https://doi.org/10.1385/1-59259-584-7:571

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-524-9

  • Online ISBN: 978-1-59259-584-6

  • eBook Packages: Springer Protocols

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