Azimzadeh Pedram, Olson John A, Balenga Nariman
University of Maryland School of Medicine, Baltimore, MD, United States.
University of Maryland School of Medicine, Baltimore, MD, United States.
Methods Cell Biol. 2017;142:89-99. doi: 10.1016/bs.mcb.2017.07.006. Epub 2017 Sep 1.
Luciferase-based assays are applied to evaluate various cellular processes due to their sensitivity and feasibility. The field of GPCR research has also benefited from this enzymatic reaction both in deorphanization campaigns and in delineation of the signaling pathways. Here, we describe the details of this assay in GPCR studies in 96-well format and will provide examples where the assay can show constitutive activity of an orphan GPCR and demonstrate the impact of cell type on the efficacy and potency of ligands.
基于荧光素酶的检测方法因其灵敏度和可行性而被应用于评估各种细胞过程。GPCR研究领域在孤儿受体鉴定活动和信号通路描绘方面也受益于这种酶促反应。在这里,我们描述了这种96孔格式的GPCR研究检测方法的细节,并将提供实例,说明该检测方法如何能够显示孤儿GPCR的组成性活性,并证明细胞类型对配体效力和效能的影响。