重组BALB/C鼠冷诱导RNA结合蛋白CIRP多克隆抗体的制备与鉴定

冷诱导RNA结合蛋白(CIRP)是第一种在哺乳动物细胞中被发现的冷休克蛋白,伴随着细胞冷应激过程过量表达.为进一步研究冷诱导RNA结合蛋白CIRP的生物学功能,构建重组BALB/C鼠CIRP的原核表达载体,重组BALB/C鼠CIRP蛋白经NI-NTA亲和树脂纯化后免疫獭兔制备抗CIRP多克隆抗体,以间接ELISA和Western blot检测抗体效价和抗原特异性.本实验成功构建了CIRP的原核表达载体,在E coli XL-1-blue得到了高效表达,利用原核表达包涵体蛋白免疫獭兔获得了高效价的CIRP多克隆抗体,该抗体能识别肝脏和睾丸中天然的CIRP蛋白.图5参14

Preparation and Identification of Recombinant Protein of CIRP'S Polyclonal Antibody in BALB/C Mice

Cold inducible RNA-binding protein (CIRP) is the first cold shock protein found in mammalian cells and overexpressed upon a temperature downshift. To further study the biological function of CIRP, polyclonal antibody of CIRP was prepared. The prokaryotic expression vector of recombination BALB / C mice CIRP was constructed, and recombinant protein CIRP was purified with the NI-NTA resin. The purified protein was used to generate polyclonal antibody in rabbits. CIRP antibody titer and antigen specificity were detected by indirect ELISA and Western blot test, respectively. The results showed that the prokaryotic expression vector of recombination BALB / C mice CIRP was constructed successfully and was highly expressed in E. coli XL-1-blue. High antibody titer polycional antibody was obtained, It could recognize natural CIRP protein in liver and testis. So, it is concluded that the high antibody titer polyclonal antibody is prepared and it can be combined with natural CIRP protein in liver and testis, which is useful for further studying the biological function of CIRP. Fig 5, Ref 14