Thermostabilization, Expression, Purification, and Crystallization of the Human Serotonin Transporter Bound to S-citalopram

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15.0K Views

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12:21 min

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November 27th, 2016

DOI :

10.3791/54792-v

November 27th, 2016

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Jonathan A. Coleman¹, Evan M. Green², Eric Gouaux¹^,³

¹Vollum Institute, Oregon Health & Science University, ²Graduate Group in Biophysics, University of California, San Francisco, ³Howard Hughes Medical Institute, Oregon Health & Science University

Podsumowanie

Rozdziały w tym wideo

0:05

Title

0:47

Transfection of Adherent HEK293S GnTI-cells for Thermostability Screen

2:24

Scintillation Proximity-based Thermostability Screen with Citalopram

4:18

Expression of the Human Serotonin Transporter in HEK293S GnTI-cells

5:24

Affinity-purification of the Serotonin Transporter for Immunization and Crystallization

7:12

Formation of Transporter-antibody Complexes and Separation by Size-exclusion Chromatography

8:21

Crystallization of Transporter-antibody Complexes by Hanging Drop

10:01

Results: Thermostabilization and Crystallization of the Serotonin Transporter in an Antidepressant-bound State

11:25

Conclusion

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