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目的制备并鉴定人端粒相关蛋白抑制和激活蛋白1(Rap1)的单克隆抗体。方法用重组原核人Rap1蛋白免疫BALB/c小鼠;细胞融合后,间接ELISA筛选阳性杂交瘤,建立分泌抗人Rap1蛋白单克隆抗体(m Ab)的杂交瘤细胞株,并进行效价测定和特异性鉴定。结果制备并获得1株稳定分泌抗人Rap1蛋白m Ab的杂交瘤细胞株。ELISA测定腹水效价高达1∶10 000以上。Western blot法、免疫沉淀和免疫荧光染色证实该m Ab能特异性识别人Rap1蛋白并可用于上述不同检测。结论成功制备获得了亲和力高、特异性好的抗人端粒相关蛋白Rap1的m Ab。
Objective To prepare and identify monoclonal antibodies against human telomere-associated protein 1 (Rap1). Methods BALB / c mice were immunized with the recombinant prokaryotic human Rap1 protein. After the cells were fused, positive hybridomas were screened by indirect ELISA. The hybridoma cell lines secreting anti-human Rap1 protein monoclonal antibody (m Ab) Specific identification. Results A hybridoma cell line stably secreting anti-human Rap1 protein m Ab was prepared and obtained. ELISA ascites titer up to 1:10 000 above. Western blot, immunoprecipitation and immunofluorescence staining confirmed that the m Ab specifically recognizes human Rap1 protein and can be used for the above different assays. Conclusion The m Ab gene of human telomere-related protein Rap1 with high affinity and specificity was obtained successfully.