Human Prolactin-Releasing Peptide Receptor Reporter Assay System

This assay utilizes proprietary human cells that have been engineered to provide constitutive expression of the Prolactin-Releasing Peptide Receptor (PrRPR).

PrRPR signals through the Gαq > PLCβ > PKC signaling pathway. This pathway leads to an increase in intracellular calcium and the concomitant activation of calcineurin, a calcium-dependent phosphatase. Ca⁺²-calcineurin, in turn, acts to dephosphorylate and activate the transcription factor NFAT3. PrRPR activation of the Ca⁺²-calcineurin > NFAT cascade is the signal transduction pathway exploited by the reporter cells provided in this kit.

INDIGO's PrRPR Reporter Cells contain an engineered luciferase reporter gene functionally linked to tandem consensus sequences of the NFAT genetic response element positioned upstream of a minimal promoter. When activated, NFAT binds to these response elements to seed the formation of a complete transcription complex that drives luciferase gene expression. Quantifying relative changes in luciferase activity in the treated reporter cells relative to the untreated cells provides a sensitive surrogate measure of druginduced changes in PrRPR 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 PrRPR, or the coupled Ca⁺²-calcineurin / NFAT signal transduction pathway.

The Reporter Cells in this kit are transiently transfected and prepared as frozen stocks using INDIGO’s 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, 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 PrRPR Reporter Cells, provided are two optimized media for use in recovering the cryopreserved cells and for diluting test samples, the reference activator PrRP-31, Luciferase Detection Reagents, and a cell culture-ready assay plate, and a detailed assay protocol. [INDIGO Catalog Nos. IB49001 | IB49001-32]

WARNING This product is not for human or veterinary use.