Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.7/614
Título: Analysis of Protein Turnover by Quantitative SNAP-Based Pulse-Chase Imaging
Autor: Bodor, Dani L.
Rodríguez, Mariluz Gómez
Moreno, Nuno
Jansen, Lars E.T.
Palavras-chave: Animals
Cell Line
Cell Survival
Fluorescent Dyes
Microscopy, Fluorescence
Recombinant Fusion Proteins
Data: 1-Jun-2012
Editora: Wiley
Citação: Bodor, 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.
Resumo: Assessment 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.
Peer review: yes
URI: http://hdl.handle.net/10400.7/614
DOI: 10.1002/0471143030.cb0808s55
Versão do Editor: http://onlinelibrary.wiley.com/doi/10.1002/0471143030.cb0808s55/abstract;jsessionid=98F7B489E8A948D98BDA41CEAD8B975B.f01t02
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