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

15,253 Views

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11:56 min

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February 12, 2014

DOI:

10.3791/51139-v

DOI:
10.3791/51139-v

•

11:56 min
February 12, 2014

•

15 Views
, , , ,
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

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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.

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

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Carrodus, N. L., Teng, K. S., Munro, K. M., Kennedy, M. J., Gunnersen, J. M. Differential Labeling of Cell-surface and Internalized Proteins after Antibody Feeding of Live Cultured Neurons. J. Vis. Exp. (84), e51139, doi:10.3791/51139 (2014).

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