Methods in molecular biology (Clifton, N.J.), 2011, Vol.769, p.403
2011
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- Autor(en) / Beteiligte
- Titel
- Measuring FRET using time-resolved FLIM
- Ist Teil von
- Methods in molecular biology (Clifton, N.J.), 2011, Vol.769, p.403
- Ort / Verlag
- United States
- Erscheinungsjahr
- 2011
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Cell migration is a process that is controlled by the formation and correct localization of protein complexes and by post-translational modification of individual proteins. Forster or fluorescent resonance energy transfer (FRET) detected using fluorescence lifetime imaging microscopy (FLIM) provides a method by which protein-protein interactions may be detected and spatially localized within a cell. This technique can be used to map protein activation states and the formation and dissolution of protein complexes that control movement of a cell. This chapter describes a protocol for detecting FRET between GFP- and mRFP1-tagged proteins in fixed adherent cells. A background to both FRET and FLIM is provided followed by an overview of the method and a full protocol for sample preparation, data acquisition, and analysis.
- Sprache
- Englisch
- Identifikatoren
- eISSN: 1940-6029DOI: 10.1007/978-1-61779-207-6_27Titel-ID: cdi_pubmed_primary_21748691
- Format
- –
- Schlagworte
- Cell Line, Tumor, Fluorescence Resonance Energy Transfer - methods, Green Fluorescent Proteins - chemistry, Green Fluorescent Proteins - metabolism, Humans, Microscopy, Fluorescence - methods, Recombinant Fusion Proteins - chemistry, Recombinant Fusion Proteins - metabolism, Software, Transfection