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Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy
JoVE 杂志
生物学
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JoVE 杂志 生物学
Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy

Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy

DOI:
10.3791/55514-v

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10:24 min

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April 09, 2017

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, , , , ,
1National Engineering Research Center of Marine Facilities Aquaculture, Marine Science College,Zhejiang Ocean University, 2Institute of Biochemistry, College of Life Sciences,Zhejiang University

Chapters

  • 00:05Title
  • 00:49Cell Seeding
  • 02:05Cell Transfection Method 1 with Liposome Formulations
  • 03:44Cell Culture on Coverslips
  • 04:37Cellular Localization Using Confocal Microscopy
  • 05:47Cellular Internalization
  • 06:42Confocal Microscopy
  • 07:43Results: Gonadotropin Releasing Hormone Receptor Internalization Detection
  • 09:21Conclusion

Summary

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This protocol describes confocal microscopy detection of G protein-coupled receptor (GPCR) internalization in mammalian cells. It includes the basic cell culture, transfection, and confocal microscopy procedure and provides an efficient and easily interpretable method to detect the subcellular localization and internalization of fusion-expressed GPCR.

Tags

GPCRLigand-activatedInternalizationConfocal MicroscopySubcellular LocalizationReceptor RecyclingHEK293 CellsTransfectionPlasmid DNAReduced Serum MediumReagentDMEMFBS

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