抗肿瘤转移相关蛋白Memo抗体的制备及组织表达谱分析

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。

Preparation of antibody against Memo protein and analysis of expression profile of Memo in mouse tissues

AIM: To prepare and characterize antibody against Memo protein and to detect the tissue distribution of Memo in mice. METHODS: Fusion protein GST-Memo was expressed and purified, and polyclonal antibody against Memo was prepared by immunizing mice. A FLAG-tagged eukaryotic expression vector pcDNA3-FLAG-Memo was constructed. The specificity of the antibody was detected by Western blot. RESULTS: An eukaryotic expression vector pcDNA3-FLAG-Memo was obtained. The polyclonal antibody was found to be specific to Memo. Memo protein was widely expressed in mouse tissues using the obtained antibody in Western blot. CONCLUSION: Antibody specific to Memo has been successfully obtained, which provides useful tool for investigation into Memo-associated mechanisms of tumor metastasis and invasiveness.