Related Experiment Video
Updated: Jan 24, 2026

The Cell-based L-Glutathione Protection Assays to Study Endocytosis and Recycling of Plasma Membrane Proteins
Published on: December 13, 2013
Monitoring MHC-II Endocytosis and Recycling Using Cell-Surface Protein Biotinylation-Based Assays
1Experimental Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Abstract:
Most, if not all, plasma membrane proteins continuously undergo endocytosis and many rapidly recycle from endosomes back to the cell surface to maintain "stable" surface expression. We now describe a biochemical assay that is suited to follow the internalization and recycling kinetics of plasma membrane proteins. This assay involves biotinylation of plasma membrane proteins using sulfo-NHS-SS-biotin, a water-soluble, NHS-ester biotinylation reagent that contains a cleavable disulfide bond that allows for reversible labeling of proteins. Biotinylation is rapid and stable, and does not transfer from cell to cell, and the small size of the biotin probe does not affect cell function.
More Related Videos
Related Concept Videos
Receptor-mediated Endocytosis
Receptor-mediated Endocytosis
Clathrin-Mediated Endocytosis of LDL
One well-characterized example of receptor-mediated endocytosis is the...
Endocytosis
Endocytosis always begins with the plasma membrane enclosing an incoming molecule to form a transport vesicle which, in some cases, can be coated with a protein called ‘clathrin.' Endocytosed material is either...
Cell-surface Signaling
The Early Endosome: Endocytosis of Transferrin
Recycling Endosomes and Transcytosis
The recycling endosome is not a single organelle but an extensively tubulated network of recycling pathways. It functions in storing molecules or transporting them across...

