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Parallel Interrogation of β-Arrestin2 Recruitment for Ligand Screening on a GPCR-Wide Scale using PRESTO-Tango Assay
Journal JoVE
Biochimie
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Journal JoVE Biochimie
Parallel Interrogation of β-Arrestin2 Recruitment for Ligand Screening on a GPCR-Wide Scale using PRESTO-Tango Assay
DOI:

09:03 min

March 10, 2020

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Chapitres

  • 00:05Introduction
  • 00:43Primary Screening: Poly-L-Lysine Plate Preparation
  • 01:25Primary Screening: Cell Culture and Plate Seeding
  • 02:22Primary Screening: DNA Plate Preparation and Transfections
  • 03:35Primary Screening: Cell Stimulation and Luminescence Reading
  • 04:26Secondary Screening: Cell Seeding and Transfections
  • 05:59Secondary Screening: Half Log Dose-Curve Drug Plate Preparation and Luminescence Reading
  • 07:16Results: Representative Primary Screening Hit Identification and Secondary Screening Dose-Curve Responses
  • 08:23Conclusion

Summary

Traduction automatique

Given that GPCRs are attractive druggable targets, GPCR ligand screening is thus indispensable for the identification of lead compounds and for deorphanization studies. Towards these efforts, we describe PRESTO-Tango, an open-source resource platform used for simultaneous profiling of transient β-arrestin2 recruitment at approximately 300 GPCRs using a TEV-based reporter assay.

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