Analysis of oxidative stress-induced protein carbonylation using fluorescent hydrazides
dc.contributor.author | Tamarit Sumalla, Jordi | |
dc.contributor.author | Hoogh, Anouk de | |
dc.contributor.author | Obis Monné, Èlia | |
dc.contributor.author | Alsina Obiols, David | |
dc.contributor.author | Cabiscol Català, Elisa | |
dc.contributor.author | Ros Salvador, Joaquim | |
dc.date.accessioned | 2016-05-18T09:59:14Z | |
dc.date.embargoEndDate | 2025-01-01 | |
dc.date.issued | 2012 | |
dc.description.abstract | Protein carbonyl detection has been commonly used to analyze the degree of damage to proteins under oxidative stress conditions. Most laboratories rely on derivatization of carbonyl groups with dinitrophenylhydrazine followed by Western blot analysis using antibodies against the dinitrophenyl moiety. This paper describes a protein carbonyl detection method based on fluorescent Bodipy, Cy3 and Cy5 hydrazides. Using this approach, Western blot and immunodetection are no longer needed, shortening the procedure and increasing accuracy. Combination of Cy3 and Cy5 hydrazides allows multiplexing analyses in a single two-dimensional gel. Derivatization with Bodipy hydrazide allows easy matching of the spots of interest and those obtained by general fluorescent protein staining methods, which facilitates excising target proteins from the gels and identifying them. This method is effective for detecting protein carbonylation in samples of proteins submitted to metal-catalyzed oxidation “in vitro” and assessing the effect of hydrogen peroxide and chronological aging on protein oxidative damage in yeast cells. | ca_ES |
dc.description.sponsorship | This work was supported by Grants BFU2010-19193 and CSD2007-00020 from the Ministerio de Ciencia e Innovacion, and SGR0677 from the Generalitat de Catalunya and Fundacio La Marato de TV3 (Spain). | ca_ES |
dc.identifier.doi | https://doi.org/10.1016/j.jprot.2012.04.046 | |
dc.identifier.idgrec | 018162 | |
dc.identifier.issn | 1874-3919 | |
dc.identifier.uri | http://hdl.handle.net/10459.1/57045 | |
dc.language.iso | eng | ca_ES |
dc.publisher | Elsevier | ca_ES |
dc.relation | MICINN/PN2008-2011/BFU2010-19193 | |
dc.relation | MIECI/PN2004-2007/CSD2007-00020 | |
dc.relation.isformatof | Reproducció del document publicat a https://doi.org/10.1016/j.jprot.2012.04.046 | ca_ES |
dc.relation.ispartof | Journal of Proteomics, 2012, vol. 75, núm. 12, p. 3778-3788 | ca_ES |
dc.rights | (c) Elsevier, 2012 | ca_ES |
dc.rights.accessRights | info:eu-repo/semantics/restrictedAccess | ca_ES |
dc.subject | Protein carbonyls | ca_ES |
dc.subject | Oxidative stress | ca_ES |
dc.subject | Yeast | ca_ES |
dc.title | Analysis of oxidative stress-induced protein carbonylation using fluorescent hydrazides | ca_ES |
dc.type | article | ca_ES |
dc.type.version | publishedVersion | ca_ES |
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