Effect of ionic strength on charcoal adsorption assays of receptor-estradiol complexes.
Effect of ionic strength on charcoal adsorption assays of receptor-estradiol complexes.
复制标题
离子强度对受体-雌二醇复合物活性炭吸附测定的影响。
DOI：
10.1210/endo-101-4-1034
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发表时间：
1977
期刊：
影响因子：
4.8
通讯作者：
J. H. Clark
中科院分区：
文献类型：
--
作者：
E. Peck;J. H. Clark
增加离子强度对活性炭吸附测定雌激素受体的效力的影响已被检查。将未成熟大鼠子宫制备的胞质溶胶暴露于不同浓度的KCl中，并通过活性炭和羟基磷灰石吸附、凝胶过滤和/或密度梯度离心法测量[3 H]雌二醇的饱和或“特异性”结合，使用过量己烯雌酚校正非饱和或“非特异性”结合。随着KCl浓度的增加，通过活性炭吸附测量的雌二醇结合位点的数量与受体从低盐（8 S）到高盐（4S）复合物的转化成比例地减少。[3 H]雌二醇从4S受体物质上的“剥离”具有时间和活性炭浓度依赖性。可以得出结论，活性炭吸附程序定量评估了[3 H]雌二醇与低盐，8 S受体物种的结合，但不是盐提取或盐处理受体的首选试验，因为受体数量可能人为估计较低。高盐条件下受体数量的低值可能是由于雌激素受体的结合位点暴露于活性炭所致。
The effect of increasing ionic strength on the efficacy of the charcoal adsorption assay for estrogen receptors has been examined. Cytosol prepared from immature rat uteri was exposed to variable concentrations of KCl and the saturable or "specific" binding of [3H]estradiol was measured by charcoal and hydroxylapatite adsorption, gel filtration, and/or density gradient centrifugation using excess diethylstilbestrol to correct for non-saturable or "non-specific" binding. As the concentration of KCl was increased, the number of estradiol binding sites as measured via charcoal adsorption decreased in proportion to the conversion of receptor from a low salt (8S) to a high salt (4S) complex. This "stripping" of [3H]estradiol from the 4S receptor species was both time- and charcoal concentration-dependent. It is concluded that the charcoal adsorption procedure quantitatively assesses the binding of [3H]estradiol to low salt, 8S receptor species but is not the assay of choice for salt-extracted or salt-treated receptors in view of the potential for artifactually low estimates of receptor number. Low values for receptor number under high salt conditions may result from exposure of the binding site of the estrogen receptor to charcoal.