Screen Quest(TM) Live Cell Vasopressin receptor 2 (V2R) cAMP Assay Service Pack

Vasopressin receptor 2 (V2R), a key GPCR, regulates water homeostasis and fluid balance by activating adenylyl cyclase and increasing intracellular cAMP upon vasopressin binding. Dysregulation of V2R signaling is linked to conditions such as diabetes insipidus, hypertension, and heart failure. Traditional methods often fail to fully capture the signaling complexity of V2R, which primarily couples to adenylyl cyclase to modulate cAMP levels. The Screen Quest(TM) Live Cell Vasopressin Receptor 2 (V2R) cAMP Assay Service Pack is specifically designed for real-time, high-throughput monitoring of intracellular cAMP changes associated with V2R 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 V2R. This assay employs cell lines transfected to express V2R along with a promiscuous G-protein Galpha16. The co-expression of Galpha16 enables V2R, which primarily signals through Gs to activate adenylyl cyclase and increase intracellular cAMP levels, while also coupling to Gq signaling to mobilize intracellular calcium. Activation of V2R by specific ligands, such as vasopressin, 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 corresponding no-wash calcium kits. The inclusion of V2R and Galpha16 co-expression ensures robust calcium signaling. This service pack provides all necessary components for precise measurement of V2R-mediated cAMP changes using FLIPR, FDSS, or equivalent fluorescence microplate readers. It is an ideal tool for studying V2R signaling pathways and evaluating potential therapeutic compounds targeting this receptor, particularly in the context of renal diseases, fluid balance disorders, and cardiovascular research.