Screen Quest(TM) Live Cell Chemokine (CC) receptor CCR5 cAMP Assay Service Pack

CCR5 or Chemokine (CC) receptor type 5 is a member of the GPCR family that plays a crucial role in immune responses and is involved in the pathogenesis of various diseases, including HIV infection, where it serves as a co-receptor for viral entry. Additionally, CCR5 has been associated with inflammatory diseases, cancer, and tissue remodeling, making it an important therapeutic target. Traditional methods for studying GPCRs often fail to capture the full spectrum of signaling mechanisms of specific GPCRs like CCR5, which couple to adenylyl cyclase activity and intracellular cAMP production. The Screen Quest(TM) Live Cell Chemokine (CC) Receptor CCR5 cAMP Assay Service Pack is specifically designed for real-time, high-throughput monitoring of intracellular cAMP changes associated with CCR5 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 CCR5. This assay employs cell lines transfected to express CCR5 along with a promiscuous G-protein Galpha16. The Galpha16 protein allows CCR5, which typically signals through the cAMP pathway, to also couple to Gq signal transduction and mobilize intracellular calcium. Activation of CCR5 by specific ligands, such as CCL3 (MIP-1alpha), CCL4 (MIP-1beta), 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 CCR5 and Galpha16 co-expression ensures robust calcium signaling for reliable assay performance. This service pack provides all necessary components for precise measurement of CCR5-mediated cAMP changes using FLIPR, FDSS, or equivalent fluorescence microplate readers. It is an ideal tool for studying non-Gq protein-coupled CCR5 activity, enabling researchers to explore CCR5 signaling pathways and evaluate potential therapeutic compounds targeting this receptor.