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A “Dual-Addition” Calcium Fluorescence Assay for the High-Throughput Screening of Recombinant G Protein-Coupled Receptors
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Journal JoVE Biochimie
A “Dual-Addition” Calcium Fluorescence Assay for the High-Throughput Screening of Recombinant G Protein-Coupled Receptors

A “Dual-Addition” Calcium Fluorescence Assay for the High-Throughput Screening of Recombinant G Protein-Coupled Receptors

DOI:
10.3791/64505-v

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08:46 min

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December 02, 2022

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, ,
1Department of Entomology,Texas A&M University, 2Department of Biochemistry and Biophysics,Texas A&M University

Chapitres

  • 00:04Introduction
  • 01:04Calcium Fluorescence Assay and Data Analysis
  • 07:40Results: High-Throughput, Intracellular Calcium Fluorescence Assay Mediated Screening of Small Molecule Libraries on Recombinant G Protein-Coupled Receptors (GPCRs)
  • 08:08Conclusion

Summary

Traduction automatique

Traduction automatique

In this work, a high-throughput, intracellular calcium fluorescence assay for 384-well plates to screen small molecule libraries on recombinant G protein-coupled receptors (GPCRs) is described. The target, the kinin receptor from the cattle fever tick, Rhipicephalus microplus, is expressed in CHO-K1 cells. This assay identifies agonists and antagonists using the same cells in one "dual-addition" assay.

Tags

Keywords: G Protein-coupled ReceptorGPCRCalcium Fluorescence AssayHigh-throughput ScreeningDual Addition AssayAgonistAntagonistCell DensityCell CulturePlate PreparationLiquid Handling

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