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Biochemistry

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JoVE Journal Biochemistry
Purification and Reconstitution of TRPV1 for Spectroscopic Analysis
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Purification and Reconstitution of TRPV1 for Spectroscopic Analysis

Article DOI: 10.3791/57796-v • 11:53 min • July 3rd, 2018
July 3rd, 2018 •
1, 2, 1,3, 1, 1
1Department of Physiology, University of Tennessee Health Science Center, 2Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, 3CuriRX, Inc.

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Summary
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This article describes specific methods to obtain biochemical quantities of detergent-solubilized TRPV1 for spectroscopic analysis. The combined protocols provide biochemical and biophysical tools that can be adapted to facilitate structural and functional studies for mammalian ion channels in a membrane-controlled environment.

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Purification Reconstitution TRPV1 Spectroscopic Analysis Transient Receptor Potential Ion Channel Family Polymodal Activation Electron Paramagnetic Resonance Double Electron Electron Resonance Epitroscopy Conformational Changes Thermal Gating Light Independent Gating Mutagenesis Primers PCR Reaction DPN1 Enzyme Transformation E.Coli Competent Cells LB Agar Plate
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