Preparation and Characterization of Polycolnal Antibody of Galectin-7 / 现代检验医学杂志

ABSTRACT

Objective To prepare the mouse anti recombinant human Galectin-7 antibody and the antibody was characterized in bladder cancer.Methods The gene coding for Galectin-7 was amplified by PCR from the cDNA of human foreskin cells and cloned into prokaryotic expression vector pET28a.Then the recombinant plasmid pET28a/Galectin-7 was transformed into E.coli BL21 (DE3)and expressed under IPTG induction.The recombinant Galectin-7 was purified through Ni2+-NT agarosegel column and the purified Galectin-7 used as imunogen to imunize the mouse.The titer and specificity of the anti-Galectin-7antibody from the mouse were analyzed by ELISA,Western blot and immunohistochemistry,respectively.Results The recombinant Galectin-7 was successfully expressed and purified,and the polyclonal ani-Galectin-7 antibody was suc-cessfully prepared.The titer of the antiserum was 1∶32 000 by ELISA.Western blot analysis showed this antiboday reacted specifically with Galectin-7.Immunohistochemistry analysis showed the antibody could recognize the native Galectin-7 in the human bladder cancer tissue.Conclusion The preparation recombinant Galectin-7 protein was as immunogen in rabbits.It was successful to produce high titer and high specificity of anti Galectin-7 polycolonal antibody.