The current state of two-dimensional electrophoresis with immobilized pH gradients

Angelika Görg; Christian Obermaier; Günther Boguth; Alois Harder; Burghardt Scheibe; Robert Wildgruber; Walter Weiss

doi:10.1002/(SICI)1522-2683(20000401)21:6<1037::AID-ELPS1037>3.0.CO;2-V

The current state of two-dimensional electrophoresis with immobilized pH gradients

Angelika Görg, Christian Obermaier, Günther Boguth, Alois Harder, Burghardt Scheibe, Robert Wildgruber, Walter Weiss

TUM Emeriti of Excellence

Technical University of Munich

Research output: Contribution to journal › Review article › peer-review

1669 Scopus citations

Abstract

The original protocol of two-dimensional electrophoresis with immobilized pH gradient (IPG-Dalt; Gorg et al., Electrophoresis 1988, 9, 531- 546) is updated. Merits and limits of different methods for sample solubilization, sample application (by cup-loading or in-gel rehydration) with respect to the pH interval used for IPG-isoelectric focusing are critically discussed. Guidelines for running conditions of analytical and micropreparative IPG-Dalt, using wide IPGs up to pH 12 for overview patterns, or narrow IPGs for zoom-in gels for optimum resolution and detection of minor components, are stated. Results with extended separation distances as well as automated procedures are demonstrated, and a comparison between protein detection by silver staining and fluorescent dyes is given. A brief trouble shooting guide is also included.

Original language	English
Pages (from-to)	1037-1053
Number of pages	17
Journal	ELECTROPHORESIS
Volume	21
Issue number	6
DOIs	https://doi.org/10.1002/(SICI)1522-2683(20000401)21:6<1037::AID-ELPS1037>3.0.CO;2-V
State	Published - 2000

Keywords

Alkaline proteins
Automation
Immobilized pH gradient
Narrow gradients
Review
Two-dimensional polyacrylamide gel electrophoresis
Wide gradients

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10.1002/(SICI)1522-2683(20000401)21:6<1037::AID-ELPS1037>3.0.CO;2-V

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title = "The current state of two-dimensional electrophoresis with immobilized pH gradients",

abstract = "The original protocol of two-dimensional electrophoresis with immobilized pH gradient (IPG-Dalt; Gorg et al., Electrophoresis 1988, 9, 531- 546) is updated. Merits and limits of different methods for sample solubilization, sample application (by cup-loading or in-gel rehydration) with respect to the pH interval used for IPG-isoelectric focusing are critically discussed. Guidelines for running conditions of analytical and micropreparative IPG-Dalt, using wide IPGs up to pH 12 for overview patterns, or narrow IPGs for zoom-in gels for optimum resolution and detection of minor components, are stated. Results with extended separation distances as well as automated procedures are demonstrated, and a comparison between protein detection by silver staining and fluorescent dyes is given. A brief trouble shooting guide is also included.",

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year = "2000",

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AU - Görg, Angelika

AU - Obermaier, Christian

AU - Boguth, Günther

AU - Harder, Alois

AU - Scheibe, Burghardt

AU - Wildgruber, Robert

AU - Weiss, Walter

PY - 2000

Y1 - 2000

N2 - The original protocol of two-dimensional electrophoresis with immobilized pH gradient (IPG-Dalt; Gorg et al., Electrophoresis 1988, 9, 531- 546) is updated. Merits and limits of different methods for sample solubilization, sample application (by cup-loading or in-gel rehydration) with respect to the pH interval used for IPG-isoelectric focusing are critically discussed. Guidelines for running conditions of analytical and micropreparative IPG-Dalt, using wide IPGs up to pH 12 for overview patterns, or narrow IPGs for zoom-in gels for optimum resolution and detection of minor components, are stated. Results with extended separation distances as well as automated procedures are demonstrated, and a comparison between protein detection by silver staining and fluorescent dyes is given. A brief trouble shooting guide is also included.

AB - The original protocol of two-dimensional electrophoresis with immobilized pH gradient (IPG-Dalt; Gorg et al., Electrophoresis 1988, 9, 531- 546) is updated. Merits and limits of different methods for sample solubilization, sample application (by cup-loading or in-gel rehydration) with respect to the pH interval used for IPG-isoelectric focusing are critically discussed. Guidelines for running conditions of analytical and micropreparative IPG-Dalt, using wide IPGs up to pH 12 for overview patterns, or narrow IPGs for zoom-in gels for optimum resolution and detection of minor components, are stated. Results with extended separation distances as well as automated procedures are demonstrated, and a comparison between protein detection by silver staining and fluorescent dyes is given. A brief trouble shooting guide is also included.

KW - Alkaline proteins

KW - Automation

KW - Immobilized pH gradient

KW - Narrow gradients

KW - Review

KW - Two-dimensional polyacrylamide gel electrophoresis

KW - Wide gradients

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ER -

The current state of two-dimensional electrophoresis with immobilized pH gradients

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Keywords

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