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Biochemistry

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JoVE Journal Biochemistry
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

Article DOI: 10.3791/58763-v • 06:38 min • February 7th, 2019
February 7th, 2019 •
1, 1, 1, 1, 1
1Gene Center and Department of Biochemistry, Center for Protein Science Munich (CIPSM), Ludwig-Maximilians-Universität München

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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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Transcription Factor-DNA Binding Affinities Competitive Titration Fluorescence Microscopy Dissociation Constants Anisotropy Measurements HiP-FA Binding Affinity Landscape Protein-protein Interactions Protein-drug Interactions Reproducibility Of Measurements Automated System DNA Oligomers Competitor DNA PCR Plate Agarose
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