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

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08:22 min

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July 17, 2018

DOI:

10.3791/58157-v

DOI:
10.3791/58157-v

•

08:22 min
July 17, 2018

•

2 Views
, , , , ,
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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Nolan, R., Alvarez, L. A., Griffiths, S. C., Elegheert, J., Siebold, C., Padilla-Parra, S. Calibration-free In Vitro Quantification of Protein Homo-oligomerization Using Commercial Instrumentation and Free, Open Source Brightness Analysis Software. J. Vis. Exp. (137), e58157, doi:10.3791/58157 (2018).

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