Comparative gel-based phosphoproteomics in response to signaling molecules

Handle URI:
http://hdl.handle.net/10754/562964
Title:
Comparative gel-based phosphoproteomics in response to signaling molecules
Authors:
Marondedze, Claudius ( 0000-0002-2113-904X ) ; Lilley, Kathryn S.; Thomas, Ludivine
Abstract:
The 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.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division; Bioscience Program; Core Labs
Publisher:
Humana Press
Journal:
Methods in Molecular Biology
Issue Date:
3-Sep-2013
DOI:
10.1007/978-1-62703-441-8-10
PubMed ID:
23681577
Type:
Article
ISSN:
10643745
ISBN:
9781627034401
Appears in Collections:
Articles; Bioscience Program; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorMarondedze, Claudiusen
dc.contributor.authorLilley, Kathryn S.en
dc.contributor.authorThomas, Ludivineen
dc.date.accessioned2015-08-03T11:17:15Zen
dc.date.available2015-08-03T11:17:15Zen
dc.date.issued2013-09-03en
dc.identifier.isbn9781627034401en
dc.identifier.issn10643745en
dc.identifier.pmid23681577en
dc.identifier.doi10.1007/978-1-62703-441-8-10en
dc.identifier.urihttp://hdl.handle.net/10754/562964en
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.en
dc.publisherHumana Pressen
dc.subject2D gel electrophoresisen
dc.subjectComparative analysisen
dc.subjectFluorescent stainen
dc.subjectIsoelectric focusingen
dc.subjectPhosphoproteomicsen
dc.subjectSDS gel electrophoresisen
dc.titleComparative gel-based phosphoproteomics in response to signaling moleculesen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.contributor.departmentBioscience Programen
dc.contributor.departmentCore Labsen
dc.identifier.journalMethods in Molecular Biologyen
dc.contributor.institutionDepartment of Biochemistry, Cambridge Centre for Proteomics, University of Cambridge, Cambridge, United Kingdomen
kaust.authorMarondedze, Claudiusen
kaust.authorThomas, Ludivineen
All Items in KAUST are protected by copyright, with all rights reserved, unless otherwise indicated.