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Biochemistry

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JoVE Journal Biochemistry
Calibration-free In Vitro Quantification of Protein Homo-oligomerization Using Commercial Instrumentation and Free, Open Source Brightness Analysis Software
 

Calibration-free In Vitro Quantification of Protein Homo-oligomerization Using Commercial Instrumentation and Free, Open Source Brightness Analysis Software

Article DOI: 10.3791/58157-v • 08:22 min • July 17th, 2018
July 17th, 2018 •
1, 1, 2, 2, 2, 1,2,3,4
1Cellular Imaging Group, Wellcome Centre Human Genetics, University of Oxford, 2Division of Structural Biology, Wellcome Centre Human Genetics, University of Oxford, 3Dynamic Structural Virology Group, Biocruces Health Research Centre, 4IKERBASQUE, Basque Foundation for Science

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This protocol describes a calibration-free approach for quantifying protein homo-oligomerization in vitro based on fluorescence fluctuation spectroscopy using commercial light scanning microscopy. The correct acquisition settings and analysis methods are shown.

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Keywords: Protein Homo-oligomerization In Vitro Quantification Calibration-free Confocal Microscopy Brightness Analysis Protein-protein Interactions Drug Screening Cancer Drugs Small-molecule Inhibitors Fusion Inhibitors Protein Dynamics Cell-cell Interactions FKBP12 His6 Tag MVenus Tag IMAC Size-exclusion Chromatography
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