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Measuring Plasma Membrane Protein Endocytic Rates by Reversible Biotinylation
JoVE Journal
Biology
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JoVE Journal Biology
Measuring Plasma Membrane Protein Endocytic Rates by Reversible Biotinylation
DOI:

11:32 min

December 23, 2009

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Chapters

  • 00:00Title
  • 00:56Introduction
  • 01:20Cell Culture and Solution Preparation
  • 03:37Biotinylation, Quenching and Internalization of Proteins
  • 07:30Protein Assay, Affinity Chromatography and SDS-Page Analysis of Proteins
  • 09:49Dopamine Transporter Internalization
  • 11:02Conclusion

Summary

Automatic Translation

Regulated endocytosis governs the cell surface expression levels of the majority of membrane proteins. Here we utilize reducible, membrane impermeant biotinylation reagents to measure the endocytic rate of the dopamine transporter (DAT), a polytopic membrane protein. The method facilitates a straightforward approach to measuring the endocytic rate of most plasma membrane proteins.

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