摘要
牛肾上腺皮质LDL受体经Triton X-100增溶,DEAE_(32)离子交换柱和LpB Sepharose亲和柱层析,在SDS-PAGE中有三条区带,分别在原点;Mr 160kD;Mr125kD处。进一步用8％SDS-PAGE纯化三个区带的蛋白质分别免疫新西兰大白兔所得的抗体,应用免疫印迹和ECL非同位素标记法可对牛肾上腺皮质和人皮肤纤维细胞膜上的LDL受体进行测定。
The LDL receptor in bovine adrenal cortex was solubilized and extracted with the addition of Triton X-100 and purified by a combination of DEAE 32 cellulose chromatography and LpB sepharose affinity chromatography. Three protein bands were seen on SDS-PAGE, located at the corresponding sites of the Mr 160kD and 125kD respectively. The three proteins were further purified with 8% SDS-PAGE and antisera were obtained from rabbits immunized with the purified proteins. All three antisera were able to react with LDL receptor from bovine adrenal cortex and human skin fibroblast as demonstrated by western immunoblot and ECL non-isotopic labelling. The experiments indicated that LDL receptor purified by DEAE 32 cellulose chromatography and LpB sepharose affinity chromatography can be used to prepare antibody. The antibody against LDL receptor from bovine adrenal cortex can also cross react with LDL receptor from the human skin fibroblast, and thus be used to determine the level of LDL receptor from both bovine adrenal cortex and human skin fibroblast.
关键词
低密度脂蛋白
受体
肾上腺皮质
LDL receptor
DEAE 32 cellulose chromatography
Affinity chromatography
SDS-PAGE
Antisera
