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[Preparation of PHF10 antibody and analysis of PHF10 expression gastric cancer tissues]
1Department of General Surgery, Shanghai Institute of Digestive Surgery, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200025, China.
Aim:
To prepare PHF10 antibody and check the expression of PHF10 protein in the tissues of gastric cancer and adjacent tissue.
Methods:
His-tagged PHF10 was expressed in E.coli BL21. Rabbit PHF10 polyclonal antiserum was generated by injecting the purified recombinant His-tagged PHF10 inclusion body as the antigen, and further separated by affinity purification. To confirm the specificity of the PHF10 antibody, transiently expressed Flag-PHF10 fusion protein was analyzed by immunoblotting with anti-flag monoclonal antibody control. The produced antibody Was used to check the expression of PHF10 protein in gastric cancer and adjacent tissues by Western blot.
Results:
Antibodies specifically binding to PHF10 could be obtained by immunization, and expression of PHF10 was significantly higher in gastric cancerous tissues comparing with adjacent normal tissues and GES-1 shows more PHF10 expression than gastric cancer cell lines with the generated antibody.
Conclusion:
The specific anti-PHF10 antibody is obtained and it could be used to detect the expression of PHF10 protein in gastric cancer cell lines and tissues, in which PHF10 is unregulated in gastric cancer.