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Immobilization of the first dimension in 2D blue native/SDS-PAGE allows the relative quantification of membrane proteomes

Standard

Immobilization of the first dimension in 2D blue native/SDS-PAGE allows the relative quantification of membrane proteomes. / Klepsch, Mirjam; Schlegel, Susan; Wickström, David; Friso, Giulia; van Wijk, Klaas J; Persson, Jan-Olov; de Gier, Jan-Willem; Wagner, Samuel.

in: METHODS, Jahrgang 46, Nr. 2, 10.2008, S. 48-53.

Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung

Harvard

Klepsch, M, Schlegel, S, Wickström, D, Friso, G, van Wijk, KJ, Persson, J-O, de Gier, J-W & Wagner, S 2008, 'Immobilization of the first dimension in 2D blue native/SDS-PAGE allows the relative quantification of membrane proteomes', METHODS, Jg. 46, Nr. 2, S. 48-53. https://doi.org/10.1016/j.ymeth.2008.06.017

APA

Klepsch, M., Schlegel, S., Wickström, D., Friso, G., van Wijk, K. J., Persson, J-O., de Gier, J-W., & Wagner, S. (2008). Immobilization of the first dimension in 2D blue native/SDS-PAGE allows the relative quantification of membrane proteomes. METHODS, 46(2), 48-53. https://doi.org/10.1016/j.ymeth.2008.06.017

Vancouver

Klepsch M, Schlegel S, Wickström D, Friso G, van Wijk KJ, Persson J-O et al. Immobilization of the first dimension in 2D blue native/SDS-PAGE allows the relative quantification of membrane proteomes. METHODS. 2008 Okt;46(2):48-53. https://doi.org/10.1016/j.ymeth.2008.06.017

Bibtex

@article{3f68550d07db42c5b312fda55a74fc50,
title = "Immobilization of the first dimension in 2D blue native/SDS-PAGE allows the relative quantification of membrane proteomes",
abstract = "In biological membranes many proteins are organized in complexes. The method of choice for the global analysis of the subunits of these complexes is two-dimensional blue native (2D BN)/SDS-PAGE. In the 1st dimension complexes are separated by BN-PAGE, and in the 2nd dimension their subunits are resolved by SDS-PAGE. In the currently available protocols the 1st dimension BN gel lanes get distorted during their transfer to the 2nd dimension separation gels. This leads to low reproducibility and high variation of 2D BN/SDS-gels, rendering them unsuitable for comparative analysis. We have developed 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, which lowers variation and greatly improves reproducibility of 2D BN/SDS-gels. 2D BN/SDS-PAGE with an immobilized 1st dimension was used for the comparative analysis of the cytoplasmic membrane proteomes of Escherichia coli cells overexpressing a membrane protein and to create a 2D BN/SDS-PAGE reference map of the E. coli cytoplasmic membrane proteome with 143 identified proteins from 165 different protein spots.",
keywords = "Electrophoresis, Gel, Two-Dimensional, Escherichia coli Proteins, Immobilized Proteins, Mass Spectrometry, Membrane Proteins, Proteome, Proteomics, Journal Article, Research Support, Non-U.S. Gov't",
author = "Mirjam Klepsch and Susan Schlegel and David Wickstr{\"o}m and Giulia Friso and {van Wijk}, {Klaas J} and Jan-Olov Persson and {de Gier}, Jan-Willem and Samuel Wagner",
year = "2008",
month = oct,
doi = "10.1016/j.ymeth.2008.06.017",
language = "English",
volume = "46",
pages = "48--53",
journal = "METHODS",
issn = "1046-2023",
publisher = "Academic Press Inc.",
number = "2",

}

RIS

TY - JOUR

T1 - Immobilization of the first dimension in 2D blue native/SDS-PAGE allows the relative quantification of membrane proteomes

AU - Klepsch, Mirjam

AU - Schlegel, Susan

AU - Wickström, David

AU - Friso, Giulia

AU - van Wijk, Klaas J

AU - Persson, Jan-Olov

AU - de Gier, Jan-Willem

AU - Wagner, Samuel

PY - 2008/10

Y1 - 2008/10

N2 - In biological membranes many proteins are organized in complexes. The method of choice for the global analysis of the subunits of these complexes is two-dimensional blue native (2D BN)/SDS-PAGE. In the 1st dimension complexes are separated by BN-PAGE, and in the 2nd dimension their subunits are resolved by SDS-PAGE. In the currently available protocols the 1st dimension BN gel lanes get distorted during their transfer to the 2nd dimension separation gels. This leads to low reproducibility and high variation of 2D BN/SDS-gels, rendering them unsuitable for comparative analysis. We have developed 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, which lowers variation and greatly improves reproducibility of 2D BN/SDS-gels. 2D BN/SDS-PAGE with an immobilized 1st dimension was used for the comparative analysis of the cytoplasmic membrane proteomes of Escherichia coli cells overexpressing a membrane protein and to create a 2D BN/SDS-PAGE reference map of the E. coli cytoplasmic membrane proteome with 143 identified proteins from 165 different protein spots.

AB - In biological membranes many proteins are organized in complexes. The method of choice for the global analysis of the subunits of these complexes is two-dimensional blue native (2D BN)/SDS-PAGE. In the 1st dimension complexes are separated by BN-PAGE, and in the 2nd dimension their subunits are resolved by SDS-PAGE. In the currently available protocols the 1st dimension BN gel lanes get distorted during their transfer to the 2nd dimension separation gels. This leads to low reproducibility and high variation of 2D BN/SDS-gels, rendering them unsuitable for comparative analysis. We have developed 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, which lowers variation and greatly improves reproducibility of 2D BN/SDS-gels. 2D BN/SDS-PAGE with an immobilized 1st dimension was used for the comparative analysis of the cytoplasmic membrane proteomes of Escherichia coli cells overexpressing a membrane protein and to create a 2D BN/SDS-PAGE reference map of the E. coli cytoplasmic membrane proteome with 143 identified proteins from 165 different protein spots.

KW - Electrophoresis, Gel, Two-Dimensional

KW - Escherichia coli Proteins

KW - Immobilized Proteins

KW - Mass Spectrometry

KW - Membrane Proteins

KW - Proteome

KW - Proteomics

KW - Journal Article

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

U2 - 10.1016/j.ymeth.2008.06.017

DO - 10.1016/j.ymeth.2008.06.017

M3 - SCORING: Journal article

C2 - 18674622

VL - 46

SP - 48

EP - 53

JO - METHODS

JF - METHODS

SN - 1046-2023

IS - 2

ER -