关键词: 釉原蛋白, 猪, 抗釉原蛋白多克隆抗体, SephadexG-200, DE-52纤维素

Abstract:

Objective　To prepare the polyclonal antibody to amelogenin.Methods　The fetal porcine dental enamelwas collected. Enamel matrix protein was extracted in 4M guanidine HCl (pH 7·4) with protease inhibitors present. Polyacrylamide gel filtration was included to isolate amelogenin from the initial dissociated extraction. The purified amelogenin conjugated with or without complete/incomplete Freund′s adjuvantwas then used to immunize the rabbits subcutaneously or intravenously. The spe- cific IgG antibody was further purified by DE-52 cellulose. The working concentration of IgG antibody was determined through ELISA test.Results　The Gel filtration showed that amelogenin components is atmolecularweights of 15 kD and 13 kD apparent- ly, which was consistent with those described before. The ELISA results showed that the working concentration for IgG was 1∶ 1000.Conclusion　The antibody prepared in this study can be used for the detection of amelogenin.

Key words: amelogenin, porcine, anti-amelogenin antibody, Sephadex G-200, DE-52 cellulose