Bio-Protocol (Aug 2013)

Epidermal Growth Factor (EGF) Receptor Endocytosis Assay in A549 Cells

  • Sabrina Rizzolio,
  • Luca Tamagnone

DOI
https://doi.org/10.21769/bioprotoc.847
Journal volume & issue
Vol. 3, no. 15

Abstract

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The following endocytosis assay has been optimized to assess EGF-stimulated EGFR endocytosis; but could be modified to assess other ligand-stimulated endocytosis of plasma membrane receptors (for which fluorochrome-conjugated ligands are available to track their receptor internalization). In brief, cells are treated with fluorescent EGF at 4 °C to allow binding to the receptor, but not internalization; then, endocytosis is allowed at 37 °C for different timepoints. For the setting up of this protocol we are really indebted to Dr. Letizia Lanzetti (Lanzetti et al., 2000).