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G-Protein-Coupled Receptors (GPCR)
Nociceptin-stimulated calcium response was measured in CHO-Ga16-NOP cells with Cal-520®, AM (Cat#21130). CHO-Ga16-NOP cells were seeded overnight at 60,000 cells/100 micro;L/well in a Costar black wall/clear bottom 96-well plate. The cells were incubated with equal volume (100 micro;L) of 10 micro;M Cal-520®, AM with 2 mM probenecid in Hanks with 20 mM Hepes buffer (HHBS) at 37°C for 1 hour. The Cal-520®, AM loading solution was replaced with HHBS and 1 mM probenecid. Nociceptin was added by FlexStation® (Molecular Devices) to achieve the final indicated concentrations.
GPCRs works in a manner where agonist binds with the receptors and that causes receptors to undergo conformational change, activating G-protein heterodimer. This leads to activation of its downstream signaling pathways. Ligand-induced ?-arrestin recruitment plays a critical role in GPCR regulation through receptor desensitization and internalization and also mediates G-protein independent signal transduction. The GPCRs works via secondary messengers such as cAMP, which is generated by Adenylyl Cyclase, a target for GPCRs, or DAG and IP3, which uses Phospholipase C, another target of GPCRs.
Study of GPCRs has so much potential uncovering lot of physiological processes inside the cells. One third of the drugs available on the market to date is based on targeting GPCRs shows the importance of GPCRs research. At AAT bioquest we offer a wide variety of the products.