论文部分内容阅读
为制备肝细胞胆管膜侧的呈极性分布单克隆抗体(mAb),并鉴定其特性,利用密度梯度离心法提取大鼠肝细胞胆管侧膜小体,测定特异酶活性。分离成功的肝细胞胆管侧膜小体免疫BALB/c小鼠,利用标准的杂交瘤技术制备单克隆抗体。利用ELISA法检测抗体分泌的效价。采用免疫组织化学方法观察其组织定位。Western blot对其识别的抗原进行分析。结果成功地获得1株特异性分布于肝细胞胆管膜侧的mAb1。免疫组织化学显示其分布在肝细胞胆管侧膜,在肾脏组织中也呈极性分布。Western blot结果显示mAb1分子量为110 kDa左右。说明成功地获得了肝细胞胆管膜侧特异性mAb1,并鉴定其作为极性分子分布于肝细胞胆管膜侧,它可能参与胆汁的分泌和代谢。
In order to prepare monoclonal antibody (mAb) with polarity distribution on the side of bile duct of hepatocyte, its characteristics were identified. Density gradient centrifugation was used to extract rat hepatocyte bile duct mesenchyma and activity of specific enzyme was measured. BALB / c mice were immunized with isolated hepatocellular bilaterally mesenchymal membranes and monoclonal antibodies were prepared using standard hybridoma techniques. Antibody secretion titers were measured by ELISA. Immunohistochemistry was used to observe the tissue location. Western blot analysis of its antigen recognition. Results A strain of mAb1 with specific distribution on the membrane of hepatocyte bile duct was successfully obtained. Immunohistochemistry showed that the distribution in the hepatocyte bile duct side membrane, also showed a polar distribution in the kidney tissue. Western blot results showed that the molecular weight of mAb1 was about 110 kDa. This indicated that mAb1 was successfully obtained from the bile duct of liver cells and identified as a polar molecule distributed on the bile duct side of hepatocytes. It may be involved in the secretion and metabolism of bile.