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

B. Sitek; B. Sipos; K. Pfeiffer; M. Grzendowski; G. Poschmann; E. Hawranke; K. Köper; G. Klöppel; H. E. Meyer; K. Stühler

doi:10.1007/s00216-008-2042-8

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

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

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4 Scopus citations

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.

Original language	English
Pages (from-to)	361-365
Number of pages	5
Journal	Analytical and Bioanalytical Chemistry
Volume	391
Issue number	1
DOIs	https://doi.org/10.1007/s00216-008-2042-8
State	Published - May 2008
Externally published	Yes

Keywords

Biomarker
Differential proteome analysis
Fluorescence dye
Large two-dimensional electrophoresis (2-DE)
Microdissection
Saturation DIGE

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10.1007/s00216-008-2042-8

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Sitek, B., Sipos, B., Pfeiffer, K., Grzendowski, M., Poschmann, G., Hawranke, E., Köper, K., Klöppel, G., Meyer, H. E., & Stühler, K. (2008). Establishment of "one-piece" large-gel 2-DE for high-resolution analysis of small amounts of sample using difference gel electrophoresis saturation labelling. Analytical and Bioanalytical Chemistry, 391(1), 361-365. https://doi.org/10.1007/s00216-008-2042-8

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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.",

keywords = "Biomarker, Differential proteome analysis, Fluorescence dye, Large two-dimensional electrophoresis (2-DE), Microdissection, Saturation DIGE",

author = "B. Sitek and B. Sipos and K. Pfeiffer and M. Grzendowski and G. Poschmann and E. Hawranke and K. K{\"o}per and G. Kl{\"o}ppel and Meyer, {H. E.} and K. St{\"u}hler",

note = "Funding Information: 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 f{\"u}r Bildung und Forschung (NGFN, FZ 031U119) and the Nordrhein Westfalen Ministerium f{\"u}r Wissenschaft und Forschung.",

year = "2008",

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doi = "10.1007/s00216-008-2042-8",

language = "English",

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Sitek, B, Sipos, B, Pfeiffer, K, Grzendowski, M, Poschmann, G, Hawranke, E, Köper, K, Klöppel, G, Meyer, HE & Stühler, K 2008, 'Establishment of "one-piece" large-gel 2-DE for high-resolution analysis of small amounts of sample using difference gel electrophoresis saturation labelling', Analytical and Bioanalytical Chemistry, vol. 391, no. 1, pp. 361-365. https://doi.org/10.1007/s00216-008-2042-8

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T1 - Establishment of "one-piece" large-gel 2-DE for high-resolution analysis of small amounts of sample using difference gel electrophoresis saturation labelling

AU - Sitek, B.

AU - Sipos, B.

AU - Pfeiffer, K.

AU - Grzendowski, M.

AU - Poschmann, G.

AU - Hawranke, E.

AU - Köper, K.

AU - Klöppel, G.

AU - Meyer, H. E.

AU - Stühler, K.

N1 - Funding Information: 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.

PY - 2008/5

Y1 - 2008/5

N2 - 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.

AB - 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.

KW - Biomarker

KW - Differential proteome analysis

KW - Fluorescence dye

KW - Large two-dimensional electrophoresis (2-DE)

KW - Microdissection

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JO - Analytical and Bioanalytical Chemistry

JF - Analytical and Bioanalytical Chemistry

IS - 1

ER -

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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Keywords

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