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High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy
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Biochemie
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JoVE Journal Biochemie
High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy

High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy

8,491 Views

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06:38 min

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February 07, 2019

DOI:

10.3791/58763-v

DOI:
10.3791/58763-v

•

06:38 min
February 07, 2019

•

9 Views
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1Gene Center and Department of Biochemistry, Center for Protein Science Munich (CIPSM),Ludwig-Maximilians-Universität München

Summary

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Here we present a novel method for determining binding affinities at equilibrium and in solution with high sensitivity on a large scale. This improves the quantitative analysis of transcription factor-DNA binding. The method is based on automated fluorescence anisotropy measurements in a controlled delivery system.

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Cite this Article

Jung, C., Schnepf, M., Bandilla, P., Unnerstall, U., Gaul, U. High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy. J. Vis. Exp. (144), e58763, doi:10.3791/58763 (2019).

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