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Neuroscience

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JoVE Journal Neuroscience
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

Article DOI: 10.3791/51139-v • 11:56 min • February 12th, 2014
February 12th, 2014 •
*1, *1, 1, 2, 1,3
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
* These authors contributed equally

Capitoli

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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 Labeling Cell-surface Proteins Internalized Proteins Antibody Feeding Live Cultured Neurons Transmembrane Receptor Specific Primary Antibody Extracellular Portion Permeabilized Cells Fixed Cells Labeled Secondary Antibody Protein Trafficking Endocytosis Overnight Blocking Step Unlabeled Secondary Antibody Fluorescently-labeled Secondary Antibody Fluorophore Subcellular Location
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