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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
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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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Calibration-free In Vitro Quantification Protein Homo-oligomerization Commercial Instrumentation Free Open Source Brightness Analysis Software Drug Screening Small-molecule Inhibitors Protein-protein Interactions Confocal Microscope Labeled Proteins Cancer Drugs Fusion Inhibitors Quantitative Information Protein Interactions Aggregations In Vitro Live Cell Protein Dynamics Cell-cell Interactions PLysS Cells PET-22b Vector Monomerized Human FKBP12 N-terminal His6 And MVenus Tags LB Agar Antibiotics Starter Culture
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