Human Fibroblast Growth Factor 2c/β-Klotho for Endocrine FGF Signaling Reporter Assay System

This assay utilizes proprietary human cells that have been engineered to provide constitutive expression of the Human Fibroblast Growth Factor Receptor 2c and β-Klotho, referred to herein as FGFR2c/β-Klotho. FGFR2c and β-Klotho are both single-pass transmembrane proteins. FGFR2c has an extracellular ligand-binding domain, transmembrane domain, and intracellular tyrosine kinase domain. It has been established that FGFR2c association with the co-receptor β-Klotho generates a scaffold that is essential for endocrine growth factor binding interactions, such as those with FGF-21 and FGF-19. Following growth factor binding, the activated tyrosine kinase activities of the FGFR initiate intracellular signaling cascades that may include RAS-MAPK, PI3-AKT, PLCγ and/or STAT pathways. For example, activation of the PLCγ pathway leads to an increase of intracellular calcium. One prominent outcome of the FGF/FGFR > PLCγ pathway is that calcineurin, a calcium-dependent phosphatase, dephosphorylates and activates the transcription factor NFAT. It is FGFR2c signal transduction via the Ca⁺² calcineurin / NFAT cascade that is exploited by the reporter cells provided in this kit.

INDIGO's FGFR2c/β-Klotho for Endocrine FGF Signaling Reporter Cells contain the luciferase reporter gene functionally linked to an NFAT-AP-1 responsive promoter. Activated NFAT binds to the response elements to initiate 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, dose-dependent surrogate measure of drug-induced changes in FGFR2c/β-Klotho activity. Accordingly, the principal application of this reporter assay is in the screening of test materials to quantify any functional interactions, either activating or inhibitory, that they may exert against FGFR2c/β-Klotho or the coupled NFAT 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 all-inclusive cell-based reporter assay system for FGFR2c/β-Klotho endocrine FGF signaling. In addition to Reporter Cells, included are an optimized culture medium for use in reviving the cryopreserved cells, a culture medium for use in preparing test sample treatments, the physiological endocrine activator FGF-21, Luciferase Detection Reagents, and a cell culture-ready assay plate. To re-iterate, this assay is primarily focused on the endocrine signaling activities of FGFR/FGFs. [INDIGO Catalog Nos. IB22101 | IB22101-32]

WARNING This product is not for human or veterinary use.