Human Cholinergic Receptor, Muscarinic 1 Activation Reporter Assay System

This assay utilizes proprietary human cells that have been engineered to provide constitutive expression of the Human Cholinergic receptor, Muscarinic 1 (CHRM1).

CHRM1 activation of the PLC pathways leads to 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 NFAT³. CHRM1 activation of the Ca⁺²∙calcineurin > NFAT cascade is the signal transduction pathway exploited by the reporter cells provided in this kit.

INDIGO's CHRM1 Reporter Cells contain an engineered luciferase reporter gene functionally linked to tandem consensus sequences of NFAT genetic response elements upstream of a minimal promoter. 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 reporter cells provides a sensitive surrogate measure of drug-induced changes in CHRM1 activity. Accordingly, the principal application of this reporter assay is in the screening of test compounds to quantify activating functional activities that they may exert against CHRM1, 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 CHRM1 Activation Reporter Cells, provided are two optimized media for use in recovering the cryopreserved cells and for diluting test samples, the reference activator Acetylcholine (chloride), Luciferase Detection Reagents, and a cell culture-ready assay plate.protocol. [INDIGO Catalog Nos. IB44001 | IB44001-32]

WARNING This product is not for human or veterinary use.