Screen Quest(TM) Live Cell Chemokine (CC) receptor CCR1 cAMP Assay Service Pack

Chemokine (CC) receptor type 1 (CCR1), a key member of the GPCR family, is implicated in various inflammatory and immune-related disorders, making it an important therapeutic target. Traditional methods for studying GPCRs often fail to address the signaling mechanisms of specific GPCRs like CCR1, which couple to adenylyl cyclase activity and intracellular cAMP production. The Screen Quest(TM) Live Cell Chemokine (CC) Receptor CCR1 cAMP Assay Service Pack is specifically designed for real-time, high-throughput monitoring of intracellular cAMP changes associated with CCR1 activation using transfected cell lines and Calbryte-520 wash-free calcium fluorescence detection methods. Unlike the conventional assays that require cell lysis, this assay preserves cellular integrity, enabling both temporal and spatial resolution of specific signaling events associated with CCR1. This assay employs cell lines that are transfected to express CCR1 along with a promiscuous G-protein Galpha16. The Galpha16 protein allows CCR1, which primarily signals through the cAMP pathway, to also couple to Gq signal transduction and mobilize intracellular calcium. Activation of CCR1 by specific ligands, such as CCL3 (MIP-1alpha) 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 CCR1 and Galpha16 co-expression ensures robust calcium signaling for reliable assay performance. The service pack provides all necessary components for precise measurement of CCR1-mediated cAMP changes using FLIPR, FDSS, or equivalent fluorescence microplate readers. It can be utilized for studying non-Gq protein-coupled CCR1 activity, offering researchers a reliable tool to explore CCR1 signaling pathways and assess potential therapeutic compounds targeting this receptor.