抗肿瘤转移相关蛋白Memo抗体的制备及组织表达谱分析

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

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。

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.