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报道了用两步法快速分离人胚肝低密度脂蛋白受体的方法。肝细胞膜用非离子型去污剂TritonX-100溶解后,经DE52纤维素离子交换层析和LDL偶联的Sepharose 4B亲和层析,将受体纯化了500倍。纯化后的低密度脂蛋白受体具有完整细胞和粗制细胞膜上受体的所有特征:(1)能与~(125)I-LDL高亲和性结合,并呈现明显的剂量效应;(2)与LDL的结合需要Ca~(2+),EDTA可阻断其结合;(3)能与抗LDL受体的单克隆抗体相互作用;(4)分子量为130kd。
Reported a rapid two-step separation of human embryonic liver low density lipoprotein receptor method. After the liver cell membrane was dissolved with the non-ionic detergent TritonX-100, the receptor was purified 500-fold by DE52 cellulose ion-exchange chromatography and LDL-coupled Sepharose 4B affinity chromatography. The purified LDL receptor has all the features of intact cells and receptors on the crude cell membrane: (1) It binds with high affinity to ~ (125) I-LDL with obvious dose-effect; (2) ) Binding to LDL requires Ca ~ (2 +), EDTA can block its binding; (3) can interact with anti-LDL receptor monoclonal antibody; (4) molecular weight of 130kd.