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RESOLFT Nanoscopy of Fixed Cells Using a Z-Domain Based Fusion Protein for Labelling.

PLoS ONE. 2015; 
IlgenPeter,GrotjohannTim,JansDaniel C,KilischMarkus,HellStefan W,JakobsSt
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Codon Optimization For expression of protein M, the amino acid sequence of the recombinant protein M TD [25] was reverse translated?in silico, optimized for the?E.?coli?codon usage and the cDNA was purchased from GenScript (GenScript, Piscataway, NJ, USA).? Get A Quote

摘要

RESOLFT super-resolution microscopy allows subdiffraction resolution imaging of living cells using low intensities of light. It relies on the light-driven switching of reversible switchable fluorescent proteins (RSFPs). So far, RESOLFT imaging was restricted to living cells, because chemical fixation typically affects the switching characteristics of RSFPs. In this study we created a fusion construct (FLASR) consisting of the RSFP rsEGFP2 and the divalent form of the antibody binding Z domain from protein A. FLASR can be used analogous to secondary antibodies in conventional immunochemistry, facilitating simple and robust sample preparation. We demonstrate RESOLFT super-resolution microscopy on chemically... More

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