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Comparative analysis of cytoplasmic membrane proteomes of Escherichia coli using 2D blue native/SDS-PAGE

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Comparative analysis of cytoplasmic membrane proteomes of Escherichia coli using 2D blue native/SDS-PAGE. / Schlegel, Susan; Klepsch, Mirjam; Wickström, David; Wagner, Samuel; de Gier, Jan-Willem.

In: Methods Mol Biol, Vol. 619, 2010, p. 257-69.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Schlegel, S, Klepsch, M, Wickström, D, Wagner, S & de Gier, J-W 2010, 'Comparative analysis of cytoplasmic membrane proteomes of Escherichia coli using 2D blue native/SDS-PAGE', Methods Mol Biol, vol. 619, pp. 257-69. https://doi.org/10.1007/978-1-60327-412-8_15

APA

Schlegel, S., Klepsch, M., Wickström, D., Wagner, S., & de Gier, J-W. (2010). Comparative analysis of cytoplasmic membrane proteomes of Escherichia coli using 2D blue native/SDS-PAGE. Methods Mol Biol, 619, 257-69. https://doi.org/10.1007/978-1-60327-412-8_15

Vancouver

Schlegel S, Klepsch M, Wickström D, Wagner S, de Gier J-W. Comparative analysis of cytoplasmic membrane proteomes of Escherichia coli using 2D blue native/SDS-PAGE. Methods Mol Biol. 2010;619:257-69. https://doi.org/10.1007/978-1-60327-412-8_15

Bibtex

@article{9e4eceb782ff479f93f8b48c9a0726e8,
title = "Comparative analysis of cytoplasmic membrane proteomes of Escherichia coli using 2D blue native/SDS-PAGE",
abstract = "Two-dimensional blue native (2D BN)/SDS-PAGE is the method of choice for the global analysis of the subunits of complexes in membrane proteomes. In the 1st dimension complexes are separated by BN-PAGE, and in the 2nd dimension their subunits are resolved by SDS-PAGE. The currently available protocols result in the distortion of the 1st dimension BN-gel lanes during their transfer to the 2nd dimension separation gels. This leads to low reproducibility and high variation of 2D BN/SDS-gels, making 2D BN/SDS-PAGE unsuitable for comparative analysis. Here, we present a 2D BN/SDS-PAGE protocol where the 1st dimension BN-gel is cast on a GelBond PAG film. Immobilization prevents distortion of BN-gel lanes when they are transferred to the 2nd dimension, which lowers variation and greatly improves reproducibility of 2D BN/SDS-gels. The use of 2D BN/SDS-PAGE with an immobilized first dimension is illustrated by the characterization of the cytoplasmic membrane proteome of Escherichia coli cells overexpressing cytochrome bo (3).",
keywords = "Cell Membrane, Electrophoresis, Gel, Two-Dimensional, Electrophoresis, Polyacrylamide Gel, Escherichia coli, Proteome, Journal Article, Research Support, Non-U.S. Gov't",
author = "Susan Schlegel and Mirjam Klepsch and David Wickstr{\"o}m and Samuel Wagner and {de Gier}, Jan-Willem",
year = "2010",
doi = "10.1007/978-1-60327-412-8_15",
language = "English",
volume = "619",
pages = "257--69",
journal = "Methods Mol Biol",
issn = "1064-3745",
publisher = "Humana Press",

}

RIS

TY - JOUR

T1 - Comparative analysis of cytoplasmic membrane proteomes of Escherichia coli using 2D blue native/SDS-PAGE

AU - Schlegel, Susan

AU - Klepsch, Mirjam

AU - Wickström, David

AU - Wagner, Samuel

AU - de Gier, Jan-Willem

PY - 2010

Y1 - 2010

N2 - Two-dimensional blue native (2D BN)/SDS-PAGE is the method of choice for the global analysis of the subunits of complexes in membrane proteomes. In the 1st dimension complexes are separated by BN-PAGE, and in the 2nd dimension their subunits are resolved by SDS-PAGE. The currently available protocols result in the distortion of the 1st dimension BN-gel lanes during their transfer to the 2nd dimension separation gels. This leads to low reproducibility and high variation of 2D BN/SDS-gels, making 2D BN/SDS-PAGE unsuitable for comparative analysis. Here, we present a 2D BN/SDS-PAGE protocol where the 1st dimension BN-gel is cast on a GelBond PAG film. Immobilization prevents distortion of BN-gel lanes when they are transferred to the 2nd dimension, which lowers variation and greatly improves reproducibility of 2D BN/SDS-gels. The use of 2D BN/SDS-PAGE with an immobilized first dimension is illustrated by the characterization of the cytoplasmic membrane proteome of Escherichia coli cells overexpressing cytochrome bo (3).

AB - Two-dimensional blue native (2D BN)/SDS-PAGE is the method of choice for the global analysis of the subunits of complexes in membrane proteomes. In the 1st dimension complexes are separated by BN-PAGE, and in the 2nd dimension their subunits are resolved by SDS-PAGE. The currently available protocols result in the distortion of the 1st dimension BN-gel lanes during their transfer to the 2nd dimension separation gels. This leads to low reproducibility and high variation of 2D BN/SDS-gels, making 2D BN/SDS-PAGE unsuitable for comparative analysis. Here, we present a 2D BN/SDS-PAGE protocol where the 1st dimension BN-gel is cast on a GelBond PAG film. Immobilization prevents distortion of BN-gel lanes when they are transferred to the 2nd dimension, which lowers variation and greatly improves reproducibility of 2D BN/SDS-gels. The use of 2D BN/SDS-PAGE with an immobilized first dimension is illustrated by the characterization of the cytoplasmic membrane proteome of Escherichia coli cells overexpressing cytochrome bo (3).

KW - Cell Membrane

KW - Electrophoresis, Gel, Two-Dimensional

KW - Electrophoresis, Polyacrylamide Gel

KW - Escherichia coli

KW - Proteome

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1007/978-1-60327-412-8_15

DO - 10.1007/978-1-60327-412-8_15

M3 - SCORING: Journal article

C2 - 20419415

VL - 619

SP - 257

EP - 269

JO - Methods Mol Biol

JF - Methods Mol Biol

SN - 1064-3745

ER -