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JoVE Journal
Biochemistry

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Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale
 

Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale

Article DOI: 10.3791/59038-v 10:50 min March 14th, 2019
March 14th, 2019

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Summary

Protein-protein interactions are critical for biological systems, and studies of the binding kinetics provide insights into the dynamics and function of protein complexes. We describe a method that quantifies the kinetic parameters of a protein complex using fluorescence resonance energy transfer and the stopped-flow technique.  

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Keywords: FRET Protein Complex Kinetics Association Dissociation COL1 CAND1 7-amino-4-methylcoumarin FlASH Cell Lysis Sonication Centrifugation Glutathione Beads Elution Thrombin Buffer A
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