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dc.contributor.authorMarondedze, Claudius
dc.contributor.authorLilley, Kathryn S.
dc.contributor.authorThomas, Ludivine
dc.date.accessioned2015-08-03T11:17:15Z
dc.date.available2015-08-03T11:17:15Z
dc.date.issued2013-09-03
dc.identifier.isbn9781627034401
dc.identifier.issn10643745
dc.identifier.pmid23681577
dc.identifier.doi10.1007/978-1-62703-441-8-10
dc.identifier.urihttp://hdl.handle.net/10754/562964
dc.description.abstractThe gel-based proteomics approach is a valuable technique for studying the characteristics of proteins. This technique has diverse applications ranging from analysis of a single protein to the study of the total cellular proteins. Further, protein quality and to some extent distribution can be first assessed by means of one-dimensional gel electrophoresis and then more informatively, for comparative analysis, using the two-dimensional gel electrophoresis technique. Here, we describe how to take advantage of the availability of fluorescent dyes to stain for a selective class of proteins on the same gel for the detection of both phospho- and total proteomes. This enables the co-detection of phosphoproteins as well as total proteins from the same gel and is accomplished by utilizing two different fluorescent stains, the ProQ-Diamond, which stains only phosphorylated proteins, and Sypro Ruby, which stains the entire subset of proteins. This workflow can be applied to gain insights into the regulatory mechanisms induced by signaling molecules such as cyclic nucleotides through the quantification and subsequent identification of responsive phospho- and total proteins. © Springer Science+Business Media New York 2013.
dc.publisherHumana Press
dc.subject2D gel electrophoresis
dc.subjectComparative analysis
dc.subjectFluorescent stain
dc.subjectIsoelectric focusing
dc.subjectPhosphoproteomics
dc.subjectSDS gel electrophoresis
dc.titleComparative gel-based phosphoproteomics in response to signaling molecules
dc.typeArticle
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Division
dc.contributor.departmentBioscience Core Lab
dc.contributor.departmentBioscience Program
dc.contributor.departmentCore Labs
dc.identifier.journalMethods in Molecular Biology
dc.contributor.institutionDepartment of Biochemistry, Cambridge Centre for Proteomics, University of Cambridge, Cambridge, United Kingdom
kaust.personMarondedze, Claudius
kaust.personThomas, Ludivine


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