Bylund David B, Toews Myron L
Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE, USA.
Methods Mol Biol. 2011;746:135-64. doi: 10.1007/978-1-61779-126-0_8.
The radioligand binding assay is a relatively simple but powerful tool for studying G protein-coupled receptors. There are three basic types of radioligand binding experiments: (1) saturation experiments from which the affinity of the radioligand for the receptor and the binding site density can be determined; (2) inhibition experiments from which the affinity of a competing, unlabeled compound for the receptor can be determined; and (3) kinetic experiments from which the forward and reverse rate constants for radioligand binding can be determined. Detailed methods for typical radioligand binding assays for G protein-coupled receptors in membranes and intact cells are presented for these types of experiments. Detailed procedures for analysis of the data obtained from these experiments are also given.
放射性配体结合测定法是研究G蛋白偶联受体的一种相对简单但强大的工具。放射性配体结合实验有三种基本类型：（1）饱和实验，通过该实验可以确定放射性配体对受体的亲和力和结合位点密度；（2）抑制实验，通过该实验可以确定竞争性未标记化合物对受体的亲和力；（3）动力学实验，通过该实验可以确定放射性配体结合的正向和反向速率常数。针对这些类型的实验，介绍了用于膜和完整细胞中G蛋白偶联受体的典型放射性配体结合测定的详细方法。还给出了对从这些实验中获得的数据进行分析的详细程序。
