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目的 :研制出能特异与p5 3结合的单克隆抗体 ,使对 p5 3的检测得到更为广泛的应用。 方法 :以原核表达的 p5 3-GST融合蛋白为免疫原 ,常规方法作细胞融合 ,间接酶联免疫吸附试验 (ELISA)双筛和免疫组织化学(IHC)筛选 ,将能稳定分泌抗人p5 3单抗的杂交瘤细胞株的细胞上清 (命名为M12 6 )与常用p5 3进口单抗PAB180 1(ZYMED公司 )作对比实验。结果 :此单抗适用于ELISA、IHC、免疫印迹及免疫沉淀等方面的研究 ,并且在一定程度上优于PAB180 1。结论 :新制备的单抗M12 6可考虑取代进口单抗PAB180 1而用于p5 3的表达及突变研究。
Objective: To develop a monoclonal antibody that specifically binds to p53, making detection of p53 more widely used. METHODS: The prokaryotic expressed p5 3-GST fusion protein was used as immunogen. Conventional methods were used for cell fusion. Indirect enzyme-linked immunosorbent assay (ELISA) double-screening and immunohistochemical (IHC) screening will stably secrete anti-human p53. The cell supernatant (named M12 6) of the monoclonal antibody hybridoma cell line was compared with the commonly used p53 imported monoclonal antibody PAB180 1 (ZYMED). RESULTS: This monoclonal antibody was suitable for ELISA, IHC, immunoblotting and immunoprecipitation, and it was better than PAB180 1 to some extent. Conclusion : The newly prepared monoclonal antibody M12 6 can be used to replace the import of monoclonal antibody PAB180 1 for expression and mutation of p53.