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Co-immunoprecipitation assays

DC Di Chen
QZ Qiaoxia Zheng
LS Long Sun
MJ Mingming Ji
YL Yan Li
HD Hongyu Deng
HZ Hong Zhang
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After transfection with the indicated plasmids and washing with PBS buffer, HeLa cells were lysed in lysis buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100 and protease inhibitor cocktail) on ice for 20 min and centrifuged at 13,000 rpm for 15 min at 4 °C. The supernatant was immunoprecipitated by GFP-Trap agarose beads (GNA-20-400, V-nanoab) for 1 h at 4 °C with rotation. After 5 washes with lysis buffer, agarose beads were collected by centrifugation (3000 rpm, 1 min) and boiled with SDS loading buffer for 10 min. Samples were subjected to immunoblotting assays.

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