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目的:制备抗Memo蛋白的特异性抗体,并鉴定其特异性,检测其在小鼠组织中的表达。方法:在大肠杆菌中表达GST-Memo融合蛋白,常规免疫小鼠制备抗体。构建了带有FLAG标签的真核表达载体pcDNA3-FLAG-Memo。用Westernblot检测抗体的特异性。结果:构建了Memo的真核表达载体pcDNA3-FLAG-Memo,制备的抗Memo抗体能特异性地识别Memo蛋白。Westernblot检测表明,Memo蛋白在不同的小鼠组织中都有一定程度的表达。结论:成功地获得抗Memo蛋白的特异性抗体并检测了其组织表达谱,为进一步研究其与恶性肿瘤的转移性、侵袭性的关系及其相关机理的研究提供了重要的制剂。
Objective: To prepare anti-Memo protein-specific antibodies, and to identify their specificity, to detect its expression in mouse tissues. Methods: The GST-Memo fusion protein was expressed in E. coli and the mice were routinely immunized to prepare the antibodies. The FLAG-tagged eukaryotic expression vector pcDNA3-FLAG-Memo was constructed. The specificity of the antibody was detected by Western blot. Results: The eukaryotic expression vector pcDNA3-FLAG-Memo of Memo was constructed and the anti-Memo antibody could specifically recognize Memo protein. Western blot analysis showed that Memo protein was expressed to a certain extent in different mouse tissues. CONCLUSIONS: The anti-Memo protein-specific antibody was successfully obtained and its tissue expression profile was detected. It provides an important preparation for further study of its relationship with metastasis and invasiveness of malignant tumor and its related mechanism.