dc.creatorSmolka, MB
dc.creatorZhou, HL
dc.creatorPurkayastha, S
dc.creatorAebersold, R
dc.date2001
dc.dateOCT 1
dc.date2014-11-13T13:03:21Z
dc.date2015-11-26T17:09:33Z
dc.date2014-11-13T13:03:21Z
dc.date2015-11-26T17:09:33Z
dc.date.accessioned2018-03-28T23:58:12Z
dc.date.available2018-03-28T23:58:12Z
dc.identifierAnalytical Biochemistry. Academic Press Inc, v. 297, n. 1, n. 25, n. 31, 2001.
dc.identifier0003-2697
dc.identifierWOS:000171406400004
dc.identifier10.1006/abio.2001.5318
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/81693
dc.identifierhttp://www.repositorio.unicamp.br/handle/REPOSIP/81693
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/81693
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1280655
dc.descriptionThe 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. (C) 2001 Academic Press.
dc.description297
dc.description1
dc.description25
dc.description31
dc.languageen
dc.publisherAcademic Press Inc
dc.publisherSan Diego
dc.publisherEUA
dc.relationAnalytical Biochemistry
dc.relationAnal. Biochem.
dc.rightsfechado
dc.sourceWeb of Science
dc.titleOptimization of the isotope-coded affinity tag-labeling procedure for quantitative proteome analysis
dc.typeArtículos de revistas


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