Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/83068
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Type: Journal article
Title: High resolution two-dimensional electrophoresis of native proteins
Author: Weiland, F.
Zammit, C.
Reith, F.
Hoffmann, P.
Citation: Electrophoresis, 2014; 35(12-13):1893-1902
Publisher: Verlag
Issue Date: 2014
ISSN: 0173-0835
1522-2683
Organisation: Institute for Photonics & Advanced Sensing (IPAS)
Statement of
Responsibility: 
Florian Weiland, Carla M. Zammit, Frank Reith and Peter Hoffmann
Abstract: Blue native PAGE (BN-PAGE) is a powerful method to separate protein complexes while preserving their native state. However, the resolution of the method is limited as complexes with similar molecular masses cannot be resolved. Here we describe native 2DE using immobilized pH-gradients in combination with BN-PAGE to resolve protein complexes by their pI and molecular mass. This method enables electrophoretic separation of proteins between pI 3 and 10 and can resolve molecular masses up to 1.2 MDa. Visualized gel spots at large molecular weight were identified using MS to confirm potential protein complexes. Several protein complexes could be identified, most prominent GroEL in complex with GroES, parts of the ribosomal machinery and membrane transport system. In summary, this method enables easy high-resolution electrophoretic separation of protein complexes.
Keywords: BN-PAGE; 2DE; IPGs; Native
Rights: © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
RMID: 0030000040
DOI: 10.1002/elps.201400060
Appears in Collections:IPAS publications

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