目的制备鼠抗DDI2蛋白单克隆抗体,并对其在细胞和组织中的定位进行初步研究。方法利用体外原核表达的DDI2蛋白,免疫BLAB/c小鼠,制备单克隆抗体。利用免疫组织化学技术明确该蛋白在组织中的定位。结果利用杂交瘤细胞技术,成功制备了两株DDI2单克隆抗体细胞系2H3、2B2。表达纯化的单克隆抗体具有较高的免疫活性。与正常对照组相比,肝损伤组织中该蛋白表达增强,且高表达于门脉周围肝实质细胞,并主要表达于细胞核。结论 DDI2的表达与四氯化碳诱导的肝损伤有关,损伤肝细胞主要表达于肝实质细胞的细胞核。
Objective To prepare DDI2 monoclonal antibody and observe distribution of DDI2 in liver tissue.Methods BLAB/c mice were immunized with DDI2 recombinant protein.Immunohistochemistry was used to observe the distribution of DDI2 in liver tissue.Results Two monoclonal antibodies(2H3 and 2B2) were prepared and purified.The monoclonal antibody showed higher immunoactivity to recombinant DDI2 protein.Immunohistochemical staining demonstrated that the DDI2 mainly scattered in nucleus of hepatocytes.Compared with the normal control tissue,DDI2 was highly expressed in the injury tissue around the portal vein area.Conclusion DDI2 mainly scattered in the nuclear of hepatocytes,and its expression may be related with hepatocytes injury induced by CCl4.