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。
To prepare and identify monoclonal antibodies distribution on rat hepatocyte canciluar membrane using ELISA and immunohistochemistry,one monoclonal antibody(MAb1) specific to hepatocyte canciluar membrane was obtained.Its location was demonstrated by Immunohistochemistry.In kindey tissues,it exhibited polar distribution.All these results showed that mAb1 might be involved in bile formation and secretion.It may participate in the mechanism of cholestasis diseases.