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Analysis of Protein Turnover by Quantitative SNAP-Based Pulse-Chase Imaging

2012-06-01Journal article Open access
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dc.contributor.authorBodor, Dani L.
dc.contributor.authorRodríguez, Mariluz Gómez
dc.contributor.authorMoreno, Nuno
dc.contributor.authorJansen, Lars E.T.
dc.date.accessioned2016-05-24T10:55:07Z
dc.date.available2016-06-01T00:30:08Z
dc.date.issued2012-06-01
dc.description.abstractAssessment of protein dynamics in living cells is crucial for understanding their biological properties and functions. The SNAP-tag, a self labeling suicide enzyme, presents a tool with unique features that can be adopted for determining protein dynamics in living cells. Here we present detailed protocols for the use of SNAP in fluorescent pulse-chase and quench-chase-pulse experiments. These time-slicing methods provide powerful tools to assay and quantify the fate and turnover rate of proteins of different ages. We cover advantages and pitfalls of SNAP-tagging in fixed- and live-cell studies and evaluate the recently developed fast-acting SNAPf variant. In addition, to facilitate the analysis of protein turnover datasets, we present an automated algorithm for spot recognition and quantification.pt_PT
dc.description.sponsorshipFCT doctoral fellowships: (SFRH/BD/74284/2010, SFRH/BD/33567/2008); FCT grants: (BIA-BCM/100557/2008, BIA-PRO/100537/2008); Fundação Calouste Gulbenkian; European Commission FP7 programme; EMBO installation grant.pt_PT
dc.identifier.citationBodor, D. L., Rodríguez, M. G., Moreno, N. and Jansen, L. E. 2012. Analysis of Protein Turnover by Quantitative SNAP-Based Pulse-Chase Imaging. Current Protocols in Cell Biology. 55:8.8:8.8.1–8.8.34.pt_PT
dc.identifier.doi10.1002/0471143030.cb0808s55pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.7/614
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherWileypt_PT
dc.relation.publisherversionhttp://onlinelibrary.wiley.com/doi/10.1002/0471143030.cb0808s55/abstract;jsessionid=98F7B489E8A948D98BDA41CEAD8B975B.f01t02pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectAnimalspt_PT
dc.subjectCell Linept_PT
dc.subjectCell Survivalpt_PT
dc.subjectCentromerept_PT
dc.subjectFluorescent Dyespt_PT
dc.subjectHumanspt_PT
dc.subjectMicroscopy, Fluorescencept_PT
dc.subjectProteinspt_PT
dc.subjectRecombinant Fusion Proteinspt_PT
dc.titleAnalysis of Protein Turnover by Quantitative SNAP-Based Pulse-Chase Imagingpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage38pt_PT
oaire.citation.startPage1pt_PT
oaire.citation.titleCurrent protocols in cell biologypt_PT
oaire.citation.volume55pt_PT
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT

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