Expression,Purification,Antibody Preparation and Preliminary Application of Tumor Protein GOLPH2

Recombinant expression of GOLPH2 and preparation of anti-GOLPH2 polyclonal antibody that may be a serologic marker of hepatocellular carcinoma(HCC) and clinical significance of the feasibility of the basis.golph2 was amplified from HepG2 cells and cloned into the prokaryotic expression vector pET21a(+)-TRX,then expressed in E.coli BL21(DE3) which was induced by isopropylthio-β-D-galactoside(IPTG).The recombinant protein GOLPH2 was purified by His-tag magnetic bead purification kit and detected by SDS-PAGE.Polyclonal antibodies was developed by immunizing BALB/c mice with purified recombinant protein,The specificity and titer of the antibody in anti-sera were determined by Western blot and ELISA respectively.The prokaryotic expression plasmid pET21a(+)-TRX-GOLPH2 was successfully constructed.The recombi-nant protein TRX-GOLPH2 could be expressed in abundance in the soluble form and got the purified pur-pose protein of 52 kD.SDS-PAGE and Western blot analysis showed that GOLPH2 protein was successfully expressed in BL21(DE3).It was showed that antiserum can specifically identify the 52 kD recombinant pro-tein,73 kD cell lysates and serum protein specifically by Western blot analysis.The polyclonal antibody a-gainst GOLPH2 protein was successfully prepared,and can be used for follow-up examination.