Häggblad J, Carlsson B, Kivelä P, Siitari H
KaroBio AB, Huddinge, Sweden.
Biotechniques. 1995 Jan;18(1):146-51.
A novel approach to direct determination of ligand binding constants for the human estrogen receptor hormone binding domain was developed. Recombinantly produced human receptor in yeast extracts was attached to scintillating microtitration plates. Radioligand binding to receptors was determined in a multi-detector scintillation counter designed for the microtitration plate format. The method was employed in equilibrium binding experiments, in binding competition tests and in determination of kinetic rate constants. The results obtained show that the methodology is valid in comparison to previously published data regarding hormone binding characteristics of estrogen receptors. Furthermore, the methodology offers several advantages over previous binding assays because the scintillating microtitration plates constitute both the binding reaction vial and the scintillant for the detection of bound radioactivity.
开发了一种直接测定人雌激素受体激素结合域配体结合常数的新方法。酵母提取物中重组产生的人受体附着在闪烁微量滴定板上。在专为微量滴定板格式设计的多探测器闪烁计数器中测定放射性配体与受体的结合。该方法用于平衡结合实验、结合竞争试验和动力学速率常数的测定。获得的结果表明，与先前发表的关于雌激素受体激素结合特性的数据相比，该方法是有效的。此外，该方法比以前的结合测定法具有几个优点，因为闪烁微量滴定板既构成结合反应瓶，又构成用于检测结合放射性的闪烁剂。
