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Mass spectrometry and co-immunoprecipitation (co-IP) assay

YC Yang Chen
YZ Yin Zhao
XY Xiaojing Yang
XR Xianyue Ren
SH Shengyan Huang
SG Sha Gong
XT Xirong Tan
JL Junyan Li
SH Shiwei He
YL Yingqin Li
XH Xiaohong Hong
QL Qian Li
CD Cong Ding
XF Xueliang Fang
JM Jun Ma
NL Na Liu
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For the IP assay, cells were lysed on ice with IP lysis buffer and sonicated. Total protein was immunoprecipitated overnight at 4 °C with 3 μg of the indicated antibodies. The immune complexes were added to PierceTM Protein A/G Magnetic Beads (Thermo Scientific) and then washed with IP Wash Buffer. The collected immune complexes were separated by SDS-PAGE and stained with Coomassie blue. Mass spectrometry was performed by Huijun Biotechnology (China). The proteins of interest in the Co-IP were detected by western blot assay. The antibodies used are listed in Supplementary Table 4.

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