Difference Gel Electrophoresis

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The Difference Gel Electrophoresis (English for 'difference gel electrophoresis') is a method for the quantity comparison of two or more samples of proteins in a 2D gel electrophoresis .

properties

The proteome can be visualized by 2D gel electrophoresis , but a comparison of two samples in two different gels is problematic due to the inhomogeneities in the preparation of the polyacrylamide gels , in the silver staining and in the extractions of the gel pieces. By marking both samples differently, they can be applied to a gel at the same time. The molecular marker is carried out by coupling of different fluorophores at Lysine or cysteines . Since lysines usually occur more frequently in proteins, it is sufficient to mark fewer lysines per protein ( minimal labeling ), while cysteines are completely marked ( saturation labeling , saturation marking ). After electrophoresis, the proteins in the gel are photographed with a fluorometer in each marked color and then individual proteins are punched out for further analysis. An identification consists of either an Edman degradation or a mass spectrometry after in-gel digestion .

Individual evidence

  1. ↑ M. Unlü, ME Morgan, JS Minden: Difference gel electrophoresis: a single gel method for detecting changes in protein extracts. In: Electrophoresis (1997), Volume 18, Issue 11, pp. 2071-2077. PMID 9420172 .
  2. ↑ MM Shaw, BM Riederer: Sample preparation for two-dimensional gel electrophoresis. In: Proteomics. Volume 3, Number 8, August 2003, pp. 1408-1417, ISSN  1615-9853 . doi : 10.1002 / pmic.200300471 . PMID 12923765 .
  3. ↑ JF Timms, R. Cramer: Difference gel electrophoresis. In: Proteomics. Volume 8, Numbers 23-24, December 2008, pp. 4886-4897, ISSN  1615-9861 . doi : 10.1002 / pmic.200800298 . PMID 19003860 .
  4. ↑ BM Riederer: Non-covalent and covalent protein labeling in two-dimensional gel electrophoresis. In: Journal of proteomics. Volume 71, Number 2, July 2008, pp. 231-244, ISSN  1874-3919 . doi : 10.1016 / j.jprot.2008.05.001 . PMID 18556257 .
  5. ^ LE McNamara, MJ Dalby, MO Riehle, R. Burchmore: Fluorescence two-dimensional difference gel electrophoresis for biomaterial applications. In: Journal of the Royal Society Interface (2010), Volume 7, Supplement 1, pp. 107-118. doi : 10.1098 / rsif.2009.0177.focus . PMID 19570793 ; PMC 2843984 (free full text).
  6. ↑ J. Minden: Comparative proteomics and difference gel electrophoresis. In: Biotechniques (2007), Volume 43, Edition 6, pp. 739, 741, 743 passim. PMID 18251249 . PDF ( Memento of the original from January 16, 2014 in the Internet Archive ) Info: The archive link was automatically inserted and not yet checked. Please check the original and archive link according to the instructions and then remove this notice. . @1@ 2Template: Webachiv / IABot / www.biotechniques.com