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Establishment of “one-piece” large-gel 2-DE for high-resolution analysis of small amounts of sample using difference gel electrophoresis saturation labelling

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Abstract

Large-gel two-dimensional gel electrophoresis (2-DE) is the method of choice for high-resolution proteome analysis of complex protein mixtures. Until now, however, the advantages of large 2-DE in combination with multiplexed fluorescence dye protein labelling has been complicated by the separate handling and analysis of the second-dimension gels. Therefore, we adapted the large 2-DE procedure allowing us to run “one-piece” large 2-DE gels (40 cm × 30 cm) in the second dimension for high resolution proteome analysis. Here, we show that in combination with fluorescence dye protein saturation labelling “one-piece” large 2-DE enables analysis of small amounts of sample (3 µg protein) for high-resolution proteome analysis.

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Abbreviations

CA:

Carrier ampholytes

PDAC:

Pancreatic ductal adenocarcinoma

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Acknowledgment

We would like to thank E. Wuttke and H.D. Knoop-Henningenfeld from the Workshop Biology, of Ruhr-University Bochum for their indispensable and skilful help in constructing the “one-piece” large gel system devices. This work was supported by Bundesministerium für Bildung und Forschung (NGFN, FZ 031U119) and the Nordrhein Westfalen Ministerium für Wissenschaft und Forschung.

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Authors and Affiliations

  1. Medizinisches Proteom-Center, Ruhr-Universität Bochum, ZKF 1.43, Universitätsstrasse 150, 44801, Bochum, Germany

    B. Sitek, K. Pfeiffer, M. Grzendowski, G. Poschmann, E. Hawranke, K. Köper, H. E. Meyer & K. Stühler

  2. Department of Pathology, Christian-Albrechts-Universität, Michaelisstr. 1, 24105, Kiel, Germany

    B. Sipos & G. Klöppel

Authors
  1. B. Sitek
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  2. B. Sipos
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  3. K. Pfeiffer
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  4. M. Grzendowski
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  5. G. Poschmann
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  6. E. Hawranke
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  7. K. Köper
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  8. G. Klöppel
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  9. H. E. Meyer
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  10. K. Stühler
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Corresponding author

Correspondence to K. Stühler.

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Sitek, B., Sipos, B., Pfeiffer, K. et al. Establishment of “one-piece” large-gel 2-DE for high-resolution analysis of small amounts of sample using difference gel electrophoresis saturation labelling. Anal Bioanal Chem 391, 361–365 (2008). https://doi.org/10.1007/s00216-008-2042-8

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  • DOI: https://doi.org/10.1007/s00216-008-2042-8

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