Strategic Screening and Characterization of the Visual GPCR-mini-G Protein Signaling Complex for Successful Crystallization

6.9K Views

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09:19 min

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March 16th, 2020

DOI :

10.3791/60747-v

March 16th, 2020

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Filip Pamula¹^,², Jonas Mühle¹, Alain Blanc³, Rony Nehmé⁴, Patricia C. Edwards⁴, Christopher G. Tate⁴, Ching-Ju Tsai¹

¹Laboratory of Biomolecular Research, Paul Scherrer Institute, ²Department of Biology, ETH Zürich, ³Center for Radiopharmaceutical Sciences, Paul Scherrer Institute, ⁴Laboratory of Molecular Biology, Medical Research Council

Summary

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Immuno affinity Agarose Resin

Retinal Binding

Protein Characterization

Bioscience Protocol

Buffer Preparation

Protein Purification

Chapters in this video

0:04

Introduction

0:45

Cell Membrane Solubilization and Protein Extraction

3:53

UV-VIS Spectroscopy

4:33

Automated Size-Exclusion Chromatography of Rhodopsin and Rhodopsin Mini-Go Complex

6:04

Deglycosylation

6:44

Results: UV-VIS Spectroscopy, SEC, and SDS-PAGE

8:34

Conclusion

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