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Differential Labeling of Cell-surface and Internalized Proteins after Antibody Feeding of Live Cultured Neurons
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Neuroscienze
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JoVE Journal Neuroscienze
Differential Labeling of Cell-surface and Internalized Proteins after Antibody Feeding of Live Cultured Neurons

Differential Labeling of Cell-surface and Internalized Proteins after Antibody Feeding of Live Cultured Neurons

DOI:
10.3791/51139-v

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11:56 min

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February 12, 2014

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, , , ,
1Department of Anatomy & Neuroscience,The University of Melbourne, 2Department of Neurobiology,Duke University Medical Center, 3Florey Institute of Neuroscience & Mental Health,The University of Melbourne

Capitoli

  • 00:05Titolo
  • 01:28Preparation of Dissociated Primary Embryonic Neuron Culture
  • 04:56Antibody Incubation with Live Neurons
  • 10:26Representative Imaging Results
  • 11:33Conclusion

Summary

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We describe a method to label protein on the surface of living neurons using a specific polyclonal antibody to extracellular epitopes. Protein bound by the antibody on the cell surface and subsequently internalized via endocytosis can be distinguished from protein remaining on, or trafficked to, the surface during the incubation.

Tags

Differential LabelingCell-surface ProteinsInternalized ProteinsAntibody FeedingLive Cultured NeuronsTransmembrane ReceptorProtein TraffickingEndocytosisFluorescent Secondary AntibodySubcellular Localization

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