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Monitoring Protein Endocytosis and Recycling Using FACS-Based Assays

Even Walseng1, Paul A Roche2

  • 1Experimental Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 1, 2019
PubMed

Cell surface MHC class II (MHC-II) is known to internalize and can recycle back to the plasma membrane from endosomes in antigen presenting cells. We now describe a simple protocol that allows one to follow the internalization kinetics of proteins from the surface of cells. Furthermore, a simple adaptation of this assay allows one to monitor the rate of appearance of internalized proteins back to the plasma membrane. This assay allows for quantitation of trafficking of proteins on even rare subsets of cells, something that is not possible with traditional biochemical assays.

Keywords:
EndocytosisFACSMHC class IIRecyclingTransmembrane proteins

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