Journal
/
Biology
/
Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
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

•

10:24 min

•

April 09, 2017

•

, , , , ,
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

Automatic Translation

Automatic Translation

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

Live Imaging of the Zebrafish Embryonic Brain by Confocal Microscopy

In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

Imaging G-protein Coupled Receptor (GPCR)-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum

Quantifying Agonist Activity at G Protein-coupled Receptors

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Characterization of G Protein-coupled Receptors by a Fluorescence-based Calcium Mobilization Assay

In vivo Clonal Tracking of Hematopoietic Stem and Progenitor Cells Marked by Five Fluorescent Proteins using Confocal and Multiphoton Microscopy

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy

Tracking Drug-induced Changes in Receptor Post-internalization Trafficking by Colocalizational Analysis

Read Article