Screen Quest(TM) Live Cell Chemokine (C-X-C motif) receptor CXCR1 cAMP Assay Service Pack

CXCR1 or C-X-C motif chemokine receptor 1, a key member of the GPCR family, plays an essential role in the immune response and is associated with inflammatory and immune disorders, including neutrophil recruitment in conditions such as asthma, chronic obstructive pulmonary disease (COPD), and cancer metastasis, making it an important therapeutic target. However, traditional methods for studying GPCRs often fail to capture the full spectrum of signaling mechanisms of specific receptors like CXCR1, which primarily couples to adenylyl cyclase and intracellular cAMP production. The Screen Quest(TM) Live Cell Chemokine (C-X-C motif) Receptor CXCR1 cAMP Assay Service Pack is specifically designed for real-time, high-throughput monitoring of intracellular cAMP changes associated with CXCR1 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 CXCR1. This assay employs cell lines transfected to express CXCR1 along with a promiscuous G-protein Galpha16. The Galpha16 protein enables CXCR1, which typically signals through the cAMP pathway, to also couple to Gq signal transduction and mobilize intracellular calcium. Activation of CXCR1 by specific ligands, such as IL-8 or Gro-alpha, 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 CXCR1 and Galpha16 co-expression ensures robust calcium signaling for reliable assay performance. This service pack provides all necessary components for precise measurement of CXCR1-mediated cAMP changes using FLIPR, FDSS, or equivalent fluorescence microplate readers. It is an ideal tool for studying non-Gq protein-coupled CXCR1 activity, enabling researchers to explore CXCR1 signaling pathways and evaluate potential therapeutic compounds targeting this receptor.