Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/194857
Full metadata record
DC FieldValueLanguage
dc.contributor.CRUESPUNIVERSIDADE DE ESTADUAL DE CAMPINASpt_BR
dc.typeArtigo de periódicopt_BR
dc.titleOptimization Of The Isotope-coded Affinity Tag-labeling Procedure For Quantitative Proteome Analysis.pt_BR
dc.contributor.authorSmolka, M Bpt_BR
dc.contributor.authorZhou, Hpt_BR
dc.contributor.authorPurkayastha, Spt_BR
dc.contributor.authorAebersold, Rpt_BR
unicamp.authorM B Smolka, Departamento de Bioquímica, Universidade Estadual de Campinas, Campinas, Sao Paulo, Brazil.pt_BR
unicamp.author.externalH Zhou,pt
unicamp.author.externalS Purkayastha,pt
unicamp.author.externalR Aebersold,pt
dc.subjectAffinity Labelspt_BR
dc.subjectCysteinept_BR
dc.subjectElectrophoresis, Gel, Two-dimensionalpt_BR
dc.subjectElectrophoresis, Polyacrylamide Gelpt_BR
dc.subjectIsotope Labelingpt_BR
dc.subjectLactalbuminpt_BR
dc.subjectMass Spectrometrypt_BR
dc.subjectMolecular Weightpt_BR
dc.subjectPeptide Fragmentspt_BR
dc.subjectProteomept_BR
dc.subjectSaccharomyces Cerevisiaept_BR
dc.subjectSaccharomyces Cerevisiae Proteinspt_BR
dc.subjectSodium Dodecyl Sulfatept_BR
dc.subjectTime Factorspt_BR
dc.subjectTrypsinpt_BR
dc.subjectUreapt_BR
dc.description.abstractThe combination of isotope coded affinity tag (ICAT) reagents and tandem mass spectrometry constitutes a new method for quantitative proteomics. It involves the site-specific, covalent labeling of proteins with isotopically normal or heavy ICAT reagents, proteolysis of the combined, labeled protein mixture, followed by the isolation and mass spectrometric analysis of the labeled peptides. The method critically depends on labeling protocols that are specific, quantitative, general, robust, and reproducible. Here we describe the systematic evaluation of important parameters of the labeling protocol and describe optimized labeling conditions. The tested factors include the ICAT reagent concentration, the influence of the protein, SDS, and urea concentrations on the labeling reaction, and the reaction time. We demonstrate that using the optimized conditions specific and quantitative labeling was achieved on standard proteins as well as in complex protein mixtures such as a yeast cell lysate.en
dc.relation.ispartofAnalytical Biochemistrypt_BR
dc.relation.ispartofabbreviationAnal. Biochem.pt_BR
dc.date.issued2001-Octpt_BR
dc.identifier.citationAnalytical Biochemistry. v. 297, n. 1, p. 25-31, 2001-Oct.pt_BR
dc.language.isoengpt_BR
dc.description.volume297pt_BR
dc.description.firstpage25-31pt_BR
dc.rightsfechadopt_BR
dc.rights.holderCopyright 2001 Academic Press.pt_BR
dc.sourcePubMedpt_BR
dc.identifier.issn0003-2697pt_BR
dc.identifier.doi10.1006/abio.2001.5318pt_BR
dc.identifier.urlhttp://www.ncbi.nlm.nih.gov/pubmed/11567524pt_BR
dc.date.available2015-11-27T12:29:00Z-
dc.date.accessioned2015-11-27T12:29:00Z-
dc.description.provenanceMade available in DSpace on 2015-11-27T12:29:00Z (GMT). No. of bitstreams: 1 pmed_11567524.pdf: 367889 bytes, checksum: 31ea4ccd63aa4aed406129d4a7ec205b (MD5) Previous issue date: 2001en
dc.identifier.urihttp://repositorio.unicamp.br/jspui/handle/REPOSIP/194857-
dc.identifier.idPubmed11567524pt_BR
Appears in Collections:Unicamp - Artigos e Outros Documentos

Files in This Item:
File SizeFormat 
pmed_11567524.pdf359.27 kBAdobe PDFView/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.