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Application of MALDI-TOF-Mass spectrometry to proteome analysis using stain-free gel electrophoresis

Application of MALDI-TOF-Mass spectrometry to proteome analysis using stain-free gel electrophoresis


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SUSNEA, Iuliana, Bogdan BERNEVIC, Michael WICKE, Li MA, Shuying LUI, Karl SCHELLANDER, Michael PRZYBYLSKI, 2013. Application of MALDI-TOF-Mass spectrometry to proteome analysis using stain-free gel electrophoresis. In: CAI, Zongwei, ed., Shuying LIU, ed.. Applications of MALDI-TOF Spectroscopy. Berlin, Heidelberg:Springer Berlin Heidelberg, pp. 37-54. ISBN 978-3-642-35664-3

@incollection{Susnea2013Appli-20075, title={Application of MALDI-TOF-Mass spectrometry to proteome analysis using stain-free gel electrophoresis}, year={2013}, doi={10.1007/128_2012_321}, number={331}, isbn={978-3-642-35664-3}, address={Berlin, Heidelberg}, publisher={Springer Berlin Heidelberg}, series={Topics in Current Chemistry}, booktitle={Applications of MALDI-TOF Spectroscopy}, pages={37--54}, editor={Cai, Zongwei and Liu, Shuying}, author={Susnea, Iuliana and Bernevic, Bogdan and Wicke, Michael and Ma, Li and Lui, Shuying and Schellander, Karl and Przybylski, Michael} }

<rdf:RDF xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:dcterms="http://purl.org/dc/terms/" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/rdf/resource/123456789/20075"> <dc:language>eng</dc:language> <dc:contributor>Przybylski, Michael</dc:contributor> <dc:creator>Lui, Shuying</dc:creator> <dc:contributor>Lui, Shuying</dc:contributor> <dcterms:abstract xml:lang="eng">The combination of MALDI-TOF-mass spectrometry with gel electrophoretic separation using protein visualization by staining procedures involving such as Coomassie Brilliant Blue has been established as a widely used approach in proteomics. Although this approach has been shown to present high detection sensitivity, drawbacks and limitations frequently arise from the significant background in the mass spectrometric analysis. In this chapter we describe an approach for the application of MALDI-MS to the mass spectrometric identification of proteins from one-dimensional (1D) and two-dimensional (2D) gel electrophoretic separation, using stain-free detection and visualization based on native protein fluorescence. Using the native fluorescence of aromatic protein amino acids with UV transmission at 343 nm as a fast gel imaging system, unstained protein spots are localized and, upon excision from gels, can be proteolytically digested and analyzed by MALDI-MS. Following the initial development and testing with standard proteins, applications of the stain-free gel electrophoretic detection approach to mass spectrometric identification of biological proteins from 2D-gel separations clearly show the feasibility and efficiency of this combination, as illustrated by a proteomics study of porcine skeleton muscle proteins. Major advantages of the stain-free gel detection approach with MALDI-MS analysis are (1) rapid analysis of proteins from 1D- and 2D-gel separation without destaining required prior to proteolytic digestion, (2) the low detection limits of proteins attained, and (3) low background in the MALDI-MS analysis.</dcterms:abstract> <dcterms:rights rdf:resource="http://nbn-resolving.org/urn:nbn:de:bsz:352-20140905103605204-4002607-1"/> <dc:contributor>Wicke, Michael</dc:contributor> <dcterms:title>Application of MALDI-TOF-Mass spectrometry to proteome analysis using stain-free gel electrophoresis</dcterms:title> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2013-03-11T10:23:14Z</dcterms:available> <dc:contributor>Susnea, Iuliana</dc:contributor> <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/20075"/> <dc:contributor>Schellander, Karl</dc:contributor> <dcterms:issued>2013</dcterms:issued> <dc:contributor>Ma, Li</dc:contributor> <dc:creator>Schellander, Karl</dc:creator> <dc:rights>deposit-license</dc:rights> <dc:creator>Susnea, Iuliana</dc:creator> <dcterms:bibliographicCitation>Applications of MALDI-TOF Spectroscopy / ed. by Zongwei Cai ... - Berlin [u.a.] : Springer, 2013. - S. 37-54. - (Topics in Current Chemistry ; 331). - ISBN 978-3-642-35664-3</dcterms:bibliographicCitation> <dc:creator>Wicke, Michael</dc:creator> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2013-03-11T10:23:14Z</dc:date> <dc:creator>Bernevic, Bogdan</dc:creator> <dc:creator>Ma, Li</dc:creator> <dc:creator>Przybylski, Michael</dc:creator> <dc:contributor>Bernevic, Bogdan</dc:contributor> </rdf:Description> </rdf:RDF>

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