小鼠DEAF1多克隆抗体的制备及应用  

The Preparation and Application of Mouse DEAF1 Polyclonal Antibody

作　　者：朱小慧[1] 盛德乔[1] 康乐[1] 伍仙凤[1] 邵文[1] 刘晓艳[1] 

机构地区：[1]三峡大学医学院,宜昌443002

出　　处：《生物技术通报》2014年第5期197-201,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(81172788);宜昌市科学技术研究与开发项目(A01301-01);湖北省高等学校优秀中青年科技创新团队计划(T201203)

摘　　要：为研究DEAF1在小鼠胰淋巴结中的内源性表达及调控外周组织抗原基因表达的机制,将DEAF1的原核表达质粒转化至E.coli BL21中,诱导获得重组DEAF1蛋白;经尿素梯度裂解包涵体纯化获得的重组DEAF1蛋白分期免疫Balb/C小鼠,获取鼠抗DEAF1的多克隆抗血清;采用ELISA、Western blot、免疫荧光、免疫沉淀分别鉴定其效价、特异性、敏感性和亲和力。结果表明,抗血清可与抗原发生特异性的免疫反应,可用于检测DEAF1的表达及荧光定位,抗血清与抗原有较高的亲和力。结果证明所制备的多克隆抗血清具有较高的特异性、敏感性和亲和力。To study endogenous expression of DEAF1 in mouse pancreatic lymph nodes and the mechanism of DEAF1 regulated expression of peripheral tissue antigen genes. The prokaryotic expression plasmid contain mouse DEAF1 cDNA was transformed into E. coli BL2I, the induced recombinant DEAF1 protein was purified by urea gradient cleavage, and was used to immunize Balb/C mouse. The anti- DEAF1 polyclonal antibody can get from mouse serum, and the specificity, sensibility, and affinity of antibody were test by ELISA, Western blot, Immunofluorescence and Immunoprecipitation. The results showed that anti-sera antibody can reacted with antigen and detected a specific band of 60 kD, and also can be used for immunofluorescence localization. Immunoprecipitation experiments showed that anti-sera antibody can bind with the antigen with high affinity. These results suggested that the DEAF1 polyclonal antiserum revealed high specificity, sensibility and affinity.

关 键 词：原核表达纯化 DEAF1 动物免疫 抗血清 抗体鉴定 

分 类 号：Q78[生物学—分子生物学]