
pmid: 26360039
Recycling of proteins such as channels, pumps, and receptors is critical for epithelial cell function. In this chapter we present a method to measure receptor recycling in polarized Madin-Darby canine kidney cells using an iodinated ligand. We describe a technique to iodinate transferrin (Tf), we discuss how (125)I-Tf can be used to label a cohort of endocytosed Tf receptor, and then we provide methods to measure the rate of recycling of the (125)I-Tf-receptor complex. We also show how this approach, which is easily adaptable to other proteins, can be used to simultaneously measure the normally small amount of (125)I-Tf transcytosis and degradation.
Cell Polarity, Receptors, Cell Surface, Endosomes, Endocytosis, Madin Darby Canine Kidney Cells, Protein Transport, Dogs, Microscopy, Fluorescence, Proteolysis, Animals
Cell Polarity, Receptors, Cell Surface, Endosomes, Endocytosis, Madin Darby Canine Kidney Cells, Protein Transport, Dogs, Microscopy, Fluorescence, Proteolysis, Animals
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