目的制备Tum5蛋白多克隆抗体并纯化。方法利用pQE30-Tum5重组质粒在大肠杆菌中通过IPTG诱导表达;亲和层析法纯化重组蛋白;以纯化的重组蛋白为抗原免疫新西兰大耳白兔制备抗Tum5多克隆抗体,ELISA方法检测抗体滴度,亲和层析法纯化兔抗Tum5多克隆抗体。结果利用pQE30-Tum5重组质粒,经原核表达和亲和层析获得Tum5蛋白;利用Tum5蛋白制备多克隆抗体,纯度大于95%。结论利用原核表达的Tum5蛋白成功地制备了兔抗Tum5多克隆抗体。
【Objective】 To prepare and purify polyclonal antibody of Tum5 protein. 【Methods】 First, we transformed the plasmid pQE30-Tum5 in E. coli and induced Tum5 protein under IPTG induction. After purification by nickel affinity chromatography, the Tum5 protein was used to raise rabbit Tum5 protein antibody. The enzyme-linked immunosorbent assay(ELISA) detected titer of rabbit Tum5 antibody. Purified polyclonal antibody of Tum5 protein. 【Results】 Tum5 protein was obtained through prokaryotic expression and affinity chromatography by Recombinant plasmid pQE30-Tum5. Polyclonal antibody was prepared by using Tum5 protein. The purity coefficient of antibody was over 95%. 【Conclusion】 Polyclonal antibody of Tum5 protein was successfully prepared by using Tum5 protein.