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TAP-MS (Tandem Affinity Purification-Mass Spectrum)

PX Pan Xie
FY Fu-Qiang Yuan
MH Ma-Sha Huang
WZ Wei Zhang
HZ Hong-Hao Zhou
XL Xi Li
ZL Zhao-Qian Liu
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The target gene expression vector with double label of Flag-strep was constructed to package lentivirus-infected target cells (TCT116), and overexpressed cell lines were Constructed, and the expression effect was detected by Western Blot. After cell lysis, the total protein was extracted, purified and eluted by streptactin resin and anti-flag antibody. The eluents of the experimental group and the control group were hydrolyzed with trypsin to obtain the polypeptide mixture of the two groups. LC-MS/MS were performed, respectively, to obtain the qualitative information of the protein. The protein interacting with the target protein can be obtained by subtracting the protein in the control group from the results of the experimental group.

Copyright and License information:  ©2021 Xie, Yuan, Huang, Zhang, Zhou, Li and Liu.
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