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Live-cell Measurement of Odorant Receptor Activation Using a Real-time cAMP Assay
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JoVE Revista Neurociencias
Live-cell Measurement of Odorant Receptor Activation Using a Real-time cAMP Assay

Live-cell Measurement of Odorant Receptor Activation Using a Real-time cAMP Assay

DOI:
10.3791/55831-v

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09:11 min

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October 02, 2017

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, , ,
1Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education,Shanghai Jiao Tong University School of Medicine, 2Department of Molecular Genetics and Microbiology,Duke University Medical Center, 3Department of Neurobiology, Duke Institute for Brain Sciences,Duke University Medical Center, 4Institute of Health Science,Chinese Academy of Science/Shanghai Jiao Tong University School of Medicine

Capítulos

  • 00:05Título
  • 00:50Plating and Transfection of Hana3A Cells
  • 03:03Transfection of Plasmids
  • 05:24Stimulation and Measuring OR Activity Using the Real-time cAMP Assay
  • 07:27Results: Kinetic Measurements of the Response of OR5AN1 to Muscone
  • 08:23Conclusion

Summary

Traducción Automática

Traducción Automática

Characterizing the function of odorant receptors serves an indispensable part in the deorphanization process. We describe a method to measure the activation of odorant receptors in real time using a cAMP assay.

Tags

Odorant ReceptorCAMP AssayLive-cell MeasurementHigh-throughput ScreeningHeterologous Expression SystemHana3A CellsTransfectionOdorant-to-receptor ActivationPhase-contrast MicroscopyCell CultureTrypsinizationCell Counting96-well PlateIncubationPlasmid ConstructsTransfection Mixture

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