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10.1038/srep28186 http://scihub22266oqcxt.onion/10.1038/srep28186 C4919659!4919659 !27339025     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=27339025 &cmd=llinks): Failed to open stream: HTTP request failed! 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Screening for protein-protein interactions using Förster resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) |pdf=|usr=}}{{27339025 }} gab.com Text Screening for protein-protein interactions using Förster resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) JO: Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=27339025 #FOAvip We present a high content multiwell plate cell-based assay approach to quantify protein interactions directly in cells using Förster resonance energy transfer (FRET) read out by automated fluorescence lifetime imaging (FLIM). Automated FLIM is implemented using wide-field time-gated detection, typically requiring only 10 s per field of view (FOV). Averaging over biological, thermal and shot noise with 100's to 1000's of FOV enables unbiased quantitative analysis with high statistical power. Plotting average donor lifetime vs. acceptor/donor intensity ratio clearly identifies protein interactions and fitting to double exponential donor decay models provides estimates of interacting population fractions that, with calibrated donor and acceptor fluorescence intensities, can yield dissociation constants. We demonstrate the application to identify binding partners of MST1 kinase and estimate interaction strength among the members of the RASSF protein family, which have important roles in apoptosis via the Hippo signalling pathway. KD values broadly agree with published biochemical measurements. Twit Text FOAVip Screening for protein-protein interactions using Förster resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) ????Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=27339025 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=27339025 #FOAvip English Wikipedia <ref>{{Cite pmid|27339025 }}</ref> Screening for protein-protein interactions using Förster resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) #MMPMID27339025 Margineanu A ; Chan JJ ; Kelly DJ ; Warren SC ; Flatters D ; Kumar S ; Katan M ; Dunsby CW ; French PM Sci Rep 2016[Jun]; 6 (ä): 28186 PMID27339025 show ga We present a high content multiwell plate cell-based assay approach to quantify protein interactions directly in cells using Förster resonance energy transfer (FRET) read out by automated fluorescence lifetime imaging (FLIM). Automated FLIM is implemented using wide-field time-gated detection, typically requiring only 10 s per field of view (FOV). Averaging over biological, thermal and shot noise with 100's to 1000's of FOV enables unbiased quantitative analysis with high statistical power. Plotting average donor lifetime vs. acceptor/donor intensity ratio clearly identifies protein interactions and fitting to double exponential donor decay models provides estimates of interacting population fractions that, with calibrated donor and acceptor fluorescence intensities, can yield dissociation constants. We demonstrate the application to identify binding partners of MST1 kinase and estimate interaction strength among the members of the RASSF protein family, which have important roles in apoptosis via the Hippo signalling pathway. KD values broadly agree with published biochemical measurements. |Animals [MESH] |COS Cells [MESH] |Chlorocebus aethiops [MESH] |Fluorescence Resonance Energy Transfer/*methods [MESH] |Green Fluorescent Proteins/metabolism [MESH] |Humans [MESH] |Intracellular Signaling Peptides and Proteins [MESH] |Isoantigens/*metabolism [MESH] |Microscopy, Fluorescence/*methods [MESH] |Optical Imaging/*methods [MESH] |Protein Interaction Domains and Motifs [MESH] |Protein Serine-Threonine Kinases/*metabolism [MESH]Screening for protein-protein interactions using Förster resonance ene(epmc).pdf DeepDyve Pubget Overpricing