Human Tropomysin Receptor Kinase C Reporter Assay System

This assay utilizes proprietary human cells that have been engineered to provide constitutive expression of Tropomysin receptor kinases C (TrkC), which is also commonly referred to as Neurotrophic Tyrosine Kinase Receptor, type 3 (NTRK3).

Neurotrophin / TrkC activation of the PLCγ pathway leads to an increase of intracellular calcium3 and the concomitant activation of calcineurin, a calcium-dependent phosphatase. Ca⁺²-calcineurin acts to dephosphorylate and activate the transcription factor NFAT. TrkC activation of the Ca⁺²∙calcineurin > NFAT cascade is the signal transduction pathway exploited by the reporter cells provided in this kit.

INDIGO's TrkC Reporter Cells contain the 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 Luc 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 TrkC 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 TrkC, 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, viability determinations or cell titer adjustments prior to assay setup. This assay kit provides the convenience of an allinclusive cell-based reporter assay system for Human TrkC. In addition to the Reporter Cells, this kit includes an optimized culture medium for use in reviving the cryopreserved cells, a culture medium for use in preparing test sample treatments, the physiological activator NT-3, Luciferase Detection Reagents, and a cell culture-ready assay plate. [INDIGO Catalog Nos. IB27031 | IB27032]

WARNING This product is not for human or veterinary use.