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GPCR Assay Kits
High-throughput GPCR screening technology for real-time measurement of intracellular changes of cAMP
GPCR Assay Kits: Real-time Measurement of cAMP in Living Cells
G Protein-Coupled Receptors (GPCRs) are a large and diverse group of membrane proteins that play a critical role in cellular communication and signal transduction. Involved in a wide range of physiological processes, GPCRs are among the most studied drug targets, with more than 30% of FDA-approved drugs targeting them.
Types of GPCRs and Their Functions:
Gi-GPCRs: Regulate adenylyl cyclase activity by inhibiting cAMP production, leading to inhibitory cellular effects.
Gs-GPCRs: Regulate adenylyl cyclase activity by stimulating cAMP production, leading to stimulatory cellular effects.
Gq-GPCRs: Activate PLC, leading to calcium release and PKC activation.
G12/13-GPCRs: Regulate the cytoskeleton and cell motility through Rho GTPases.
ACTOne GPCR Assay
ACTOne™ is a high-throughput GPCR screening technology that measures intracellular changes of the secondary messenger cyclic AMP (cAMP) in living cells, in real-time. The assay uses a proprietary modified cyclic nucleotide-gated (CNG) channel, co-localized with adenylate cyclase at the plasma membrane, as a biosensor of cAMP activity. When cAMP levels increase, the CNG channel opens, resulting in ion flux and cell membrane depolarization.
Key Features:
Real-Time Measurement: Quantifies intracellular cAMP changes using fluorescent ion indicators or membrane potential (MP) dyes. Allows flexibility in dynamic control of expression.
High Sensitivity: Detects both strong and weak GPCR activities with minimal artifacts.
Versatile Applications: Suitable for detecting endogenous GPCR responses and activities not reported in the literature. Directly measures GPCR activation. Provides both kinetic and endpoint readouts. Greater content and statistically relevant data.
Easy to use by mxing the AAVs with cells.
Faster setup and more time efficient.
Cost-effective: $498 for one plate.
Kit Components:
1. AAV-CNG/GPCR enables transient expression of both CNG and GPCRs. AAV-CNG/GPCR is a mixture of AAV-CNG and AAV-GPCR. Both viruses use CMV as promoter, which is known for driving strong and rapid expression of transgenes in a wide variety of cell types. The ratio of two viruses are optimized in each kit.
2. ACTOne MP Dye allows for real-time, sensitive detection of GPCR functionality.
Workflow:
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