Mdp3抗体的制备和应用  

The Preparation and Application of Mdp3 Antibody

作　　者：孙晓鸥[1] 塔拉[1] 周军[1] 

机构地区：[1]南开大学遗传学和细胞生物学系,天津300071

出　　处：《南开大学学报（自然科学版）》2012年第5期76-81,共6页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：国家自然科学基金(30825022)

摘　　要：Mdp3是1个由876个氨基酸组成的蛋白质,其基因位于Xq26.3.利用pET32a(+)载体,构建了原核表达His-Mdp3融合蛋白的质粒pET32a(+)-Mdp3,转化到E.coli BL21(DE3)中,用IPTG诱导融合蛋白表达.摸索得到了His-Mdp3融合蛋白的最佳表达条件,并纯化得到大量纯度较高的蛋白质.利用从细菌中纯化的His-Mdp3蛋白免疫小鼠,得到针对Mdp3的抗体,并通过免疫印迹实验验证了该抗体的特异性.最后,用该抗体对细胞进行免疫荧光显微分析,发现Mdp3在细胞内呈现特异的定位模式.Mdp3, （gene located on Xq26. 3） encodes a 876-amino-acid protein pET32a （＋）-Mdp3 recombinant plasmid was constructed and transformed into E. coli BL21 （DE3）. Then the expression condition of His-Mdp3 fusion protein was optimized, and a large amount of protein with high purity was obtained. In addition, specific antibody against Mdp3 by immunizing mice with purified protein was acquired, and its specificity by immunoblotting was examined. Immunofluorescence analysis was also performed with the antibody, and the specific cellular localization of Mdp3 was revealed.

关 键 词：Mdp3 蛋白纯化 抗体 免疫荧光 

分 类 号：Q813[生物学—生物工程]