Isolation of EGFR-containing early endosomes.

Clathrin‐mediated EGFR endocytosis as a potential therapeutic strategy for overcoming primary resistance of EGFR TKI in wild‐type EGFR non‐small cell lung cancer

“…To isolate EGFR in early endosomes, immunoprecipitation was conducted as previously described. 28 For immunoprecipitation, 1 mg of protein lysate was incubated with Protein A/G agarose beads (Thermo Fisher Scientific) and anti-EEA1 antibody for 10 h. The beads were then washed thoroughly three times with lysis buffer, and the proteins were extracted in sample buffer by boiling for 10 min. Western blot assays were then performed as described above.…”