目的制备特异性识别CUE结构域蛋白2(CUE domain containing 2,CUEDC2)的单克隆抗体(mAb),并检测CUEDC2蛋白在小鼠组织中的表达。方法在大肠杆菌中融合表达GST—hCUEDC2蛋白,纯化后免疫小鼠,制备mAb。采用Western blot方法检测mAb的特异性以及小鼠组织中CUEDC2蛋白的表达。结果获得了可同时识别人源和鼠源CUEDC2mAb,利用此mAb检测小鼠组织CUEDC2蛋白的表达水平,发现该蛋白的表达在不同小鼠组织中存在差异。结论’制备的CUEDC2 mAb特异性好,并可检测CUEDC2蛋白在小鼠组织中的表达,该mAb的制备为进一步研究CUEDC2的功能奠定了基础。
Objective To prepare monoclonal antibody (mAb) against CUE domain containing 2 (CUEDC2) protein and detect the expression level of CUEDC2 in mouse tissues. Methods Fusion protein GST-human CUEDC2 (hCUEDC2) was expressed and purified, and mAb against CUEDC2 was prepared by using hybridoma technique. The specificity of the antibody and CUEDC2 expression in mouse tissues were detected by Western blot analysis. Results We obtained mAb against CUEDC2 protein, which recognized specifically both hCUEDC2 and mouse CUEDC2. The mAb could be used to detect expression level of CUEDC2 protein, which was different in various mouse tissues. Conclusion The prepared CUEDC2 has good mAb specificity and can be used to detect expression of CUEDC2 protein in mouse tissue, thus providing a useful tool for further investigation into the functions of CUEDC2.