Zafar A, Thampan R V
School of Life Sciences, University of Hyderabad, India.
Protein Expr Purif. 1993 Dec;4(6):534-8. doi: 10.1006/prep.1993.1070.
A relatively inexpensive yet highly efficient and extremely rapid procedure has been developed for the isolation and purification of estrogen receptor from the goat uterine cytosol. Greater than 1 mg of purified receptor protein could be obtained from 75 g of uterine tissue within a period of < 24 h, following this protocol. The procedure does not require the use of an ultracentrifuge, a cold room, or a column chromatography setup. The entire isolation procedure, which involves adsorption to and selective elution from different chromatography matrices, is carried out at the laboratory bench using beakers kept in an ice bath. Antibodies raised against this receptor cross-react with the goat uterine estrogen receptor activation factor, a DNA binding protein having no capacity to bind estradiol but which dimerizes with the nonactivated estrogen receptor, an estrogen receptor incapable of binding to DNA on its own.
已开发出一种相对廉价但高效且极其快速的方法，用于从山羊子宫胞质溶胶中分离和纯化雌激素受体。按照此方案，在不到24小时的时间内，从75克子宫组织中可获得超过1毫克的纯化受体蛋白。该方法不需要使用超速离心机、冷藏室或柱色谱装置。整个分离过程包括吸附到不同的色谱基质上并从其选择性洗脱，在实验室工作台上使用置于冰浴中的烧杯进行。针对该受体产生的抗体与山羊子宫雌激素受体激活因子发生交叉反应，该激活因子是一种DNA结合蛋白，无结合雌二醇的能力，但可与未激活的雌激素受体二聚化，而未激活的雌激素受体自身无结合DNA的能力。
