Screen Quest(TM) Live Cell Chemokine (CC) receptor CCR3 cAMP Assay Service Pack

CCR3 or Chemokine (CC) receptor type 3, a key member of the GPCR family, is critically involved in various inflammatory and allergic conditions, including asthma, eosinophilic disorders, and certain autoimmune diseases, making it an important therapeutic target. Traditional methods for studying GPCRs often fail to capture the signaling mechanisms of specific GPCRs like CCR3, which couple to adenylyl cyclase activity, intracellular cAMP production and calcium flux. The Screen Quest(TM) Live Cell Chemokine (CC) Receptor CCR3 cAMP Assay Service Pack is specifically designed for real-time, high-throughput monitoring of intracellular cAMP changes associated with CCR3 activation using transfected cell lines and Calbryte(TM) 520 wash-free calcium fluorescence detection methods. Unlike conventional assays requiring cell lysis, this assay preserves cellular integrity, enabling both temporal and spatial resolution of specific signaling events associated with CCR3. This assay employs cell lines transfected to express CCR3 along with a promiscuous G-protein Galpha16. The Galpha16 protein allows CCR3, which primarily signals through the cAMP pathway, to also couple to Gq signal transduction and mobilize intracellular calcium. Activation of CCR3 by specific ligands, such as CCL11 (Eotaxin-1), CCL24 (Eotaxin-2), or CCL5 (RANTES), can be detected using calcium-sensitive dyes like 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 CCR3 and Galpha16 co-expression ensures robust calcium signaling for reliable assay performance. This service pack provides all necessary components for precise measurement of CCR3-mediated cAMP changes using FLIPR, FDSS, or equivalent fluorescence microplate readers. It is an ideal tool for studying coupled CCR3 activity, enabling researchers to explore CCR3 signaling pathways and evaluate potential therapeutic compounds targeting this receptor.