Adaptation of aequorin functional assay to high throughput screening.

摘要: AequoScreen™, a cellular aequorin-based functional assay, has been optimized for luminescent high-throughput screening (HTS) of G protein-coupled receptor (GPCRs). AequoScreen is homogeneous assay in which the cells are loaded with apoaequorin cofactor coelenterazine, diluted buffer, and injected into plates containing samples to be tested. A flash light emitted following calcium increase resulting from activation GPCR by sample. Here we have validated new plate reader, Hamamatsu Photonics FDSS6000, HTS 96- 384-well CHO-K1 stably coexpressing mitochondrial different GPCRs (AequoScreen cell lines). The acquisition time, type, number per well obtain concentration-response curves 4000 cells/well high signal: background ratio. FDSS6000 lines allow reading twenty or 1 h Z’ values 0.71 0.7...