Author:
Date:
2013-08-05
[Abstract] 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).
[摘要] 以下内吞测定已经被优化以评估EGF刺激的EGFR内吞作用; 但可以修改以评估其他配体刺激的细胞膜受体的内吞作用(其中荧光染料偶联的配体可用于追踪它们的受体内化)。 简言之,在4℃下用荧光EGF处理细胞以允许结合受体,但不内化; 然后,在37℃下对不同的时间点进行内吞作用。 为了建立该方案,我们真正感谢Letizia Lanzetti博士(Lanzetti等人,2000)。
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