Human Arginine Vasopressin Receptor 1B Reporter Assay System

This assay utilizes proprietary human cells that have been engineered to provide constitutive expression of the Human Arginine Vasopressin Receptor 1B (AVPR1B).

Upon activation by its physiological ligand vasopressin, AVPR1B activates Gα_q/11 to mediate several downstream pathways. This includes activation of the phospholipase C (PLC) pathway, which leads to the release of inositol triphosphate (IP3), resulting in an increase in intracellular calcium and the concomitant activation of calcineurin, a calcium dependent phosphatase. Ca⁺²∙calcineurin acts to dephosphorylate and activate the transcription factor NFAT¹². The AVPR1B signal transduction cascade that leads to activation of Ca⁺²∙calcineurin > NFAT is the pathway exploited by the reporter cells provided in this kit.

INDIGO's AVPR1B Reporter Cells contain an engineered luciferase reporter gene functionally linked to tandem consensus sequences of the NFAT genetic response elements and a minimal promoter. Activated NFAT binds to these response elements to seed the formation of a complete transcription complex that drives luciferase reporter gene expression. Quantifying relative changes in luciferase activity in the treated reporter cells relative to the untreated cells provides a sensitive surrogate measure of drug-induced changes in AVPR1B activity.

The principal application of this reporter assay is in the screening of test samples to quantify functional interactions, either activating or inhibitory, that they may exert against AVPR1B, or the coupled Ca⁺² ∙ calcineurin / NFAT signal transduction pathway.

INDIGO’s Reporter Cells are transiently transfected and prepared as frozen stocks using a proprietary CryoMite™ process. This cryo-preservation method allows for the immediate dispensing of healthy, division-competent reporter cells into assay plates. There is no need for intermediate treatment steps such as spin-and-rinse of cells, extended pre-culture, viability determinations, or cell titer adjustments prior to assay setup.

INDIGO's assay kits provide the convenience of an all-inclusive cell-based assay system. In addition to AVPR1B Reporter Cells, provided are two optimized media for use in recovering the cryopreserved cells and for diluting test samples, the reference activator Vasopressin, Luciferase Detection Reagents, and a cell culture-ready assay plate. [INDIGO Catalog Nos. IB37101 | IB37102]

WARNING This product is not for human or veterinary use.