AAV ACTOne Gs-GPCR Assay Kit-VIPR2

The AAV ACTOne GPCR Assay Kits are live-cell-based cAMP biosensor systems designed for rapid and high-throughput functional screening of G-protein coupled receptors (GPCRs) by combination AAV. Kits are available for both Gi-GPCR and Gs-GPCR assays.

AAV ACTOne Gi-GPCR Assay Kits:

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AAV ACTOne Gs-GPCR Assay Kits:

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Advantages of AAV ACTOne GPCR Assay Kits:

• Combined ACTOne and AAV Technologies
• Transient Expression of CNG and GPCR
• Real Time Assay on Living Cell
• High Sensitivity and Big assay window
• Both Kinetic and Endpoint Assay
• Easy to use by mixing the AAVs with cells
• Faster setup and time efficiency
  
• Reduced risk of genetic drift, 
• Simplified workflows, 
• flexibility in dynamic control of expression 
• Low Cost: $498 for one plate

ACTOne GPCR Assay

ACTOne™ is the only high-throughput GPCR screening technology that can directly measure the intracellular changes of the secondary messenger cyclic AMP (cAMP) in living cells, in real-time. It uses a proprietary modified cyclic nucleotide-gated (CNG) channel, which is co-localized with adenylate cyclase at the plasma membrane, as a biosensor of cAMP activity. The CNG channel opens when the cAMP level near the plasma membrane increases, resulting in ion flux and cell membrane depolarization. The influx of cations through the CNG channel can be quantified using fluorescent ion indicators or membrane potential (MP) dyes. It provides information on real time intracellular cAMP changes and is highly sensitive. By combining kinetic and endpoint readouts, we are able to capture and analyze transient responses from endogenous GPCRs and weak responses caused by weak Gs or Gi coupled GPCR activities. Using ACTOne, we are able to detect the subcellular cAMP concentration changes directly caused by GPCR activation. Real-time kinetic readouts minimize artifacts, and provide greater content and more statistically relevant data. The intensity of signal increase caused by GPCR activation is directly related to the receptor number on cell surface. Using ACTOne assay, we were able to detect activities of some endogenous Gs coupled receptors in HEK293 cells that have not been reported in literature. In addition, we have also detected weak Gs coupled activity of a GPCR that was widely considered to be only linked to Gq coupled pathway.  The ACTOne assay also provides a useful tool for GPCR de-orphanization.