Screen Quest(TM) Live Cell Opioid-Like receptor hKOR cAMP Assay Service Pack

hKOR, also known as the Human Kappa Opioid Receptor, is a key member of the G-protein coupled receptor (GPCR) family, involved in various neurological and psychiatric conditions, including pain management, depression, and addiction. Traditional methods for studying GPCRs often fail to address the full spectrum of signaling mechanisms through specific receptors like hKOR, which primarily couples to Gi/o-mediated inhibition of adenylyl cyclase and reduction in intracellular cAMP. The Screen Quest(TM) Live Cell Opioid-Like Receptor hKOR cAMP Assay Service Pack is specifically designed for real-time, high-throughput monitoring of intracellular cAMP changes associated with hKOR activation using transfected cell lines and Calbryte(TM) 520 wash-free calcium fluorescence detection methods. Unlike conventional assays that require cell lysis, this assay preserves cellular integrity, enabling both temporal and spatial resolution of specific signaling events associated with hKOR. This assay employs cell lines transfected to express hKOR alongside a promiscuous G-protein Galpha16. The co-expression of Galpha16 enables hKOR, which normally signals via Gi/o-mediated inhibition of adenylyl cyclase and reduction in intracellular cAMP, to also couple to Gq signaling, leading to calcium flux. Activation of hKOR by specific ligands, such as dynorphins or salvinorin A, can be detected using calcium-sensitive dyes such as Calbryte(TM) 520 AM, Cal-520(R) AM, Fluo-8(R) AM, Fluo-4(TM) AM, or other corresponding no-wash calcium kits. This service pack provides all necessary components for precise measurement of hKOR-mediated cAMP changes using FLIPR, FDSS, or equivalent fluorescence microplate readers. It enables researchers to explore hKOR signaling pathways and evaluate potential therapeutic compounds targeting this receptor, particularly in the context of pain relief, mood disorders, and addiction research.