Mouse Fibroblast Growth Factor 1c/β-Klotho (mFGFR1c/β-Klotho) for Endocrine FGF Signaling Reporter Assay System

INDIGO's Mouse Fibroblast Growth Factor 1c/β-Klotho (mFGFR1c/β-Klotho) for Endocrine FGF Signaling Reporter Assay System utilizes proprietary cells that have been engineered to provide constitutive expression of the Mouse Fibroblast Growth Factor Receptor 1c and Mouse β-Klotho, referred to herein as mFGFR1c/β-Klotho. mFGFR1c and β-Klotho are both single-pass transmembrane proteins. mFGFR1c has an extracellular ligand-binding domain, transmembrane domain, and intracellular tyrosine kinase domain. It has been established that mFGFR1c association with the co-receptor β-Klotho generates a scaffold that is essential for endocrine growth factor binding interactions, such as those with FGF-19 and FGF-21. 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 mFGFR1c/β-Klotho signal transduction via the Ca⁺²∙calcineurin / NFAT cascade that is exploited by the reporter cells provided in this kit.

INDIGO's mFGFR1c/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 mFGFR1c/βKlotho 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 mFGFR1c/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 mFGFR1c/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 Murine FGF-21 (mFGF-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. M22001, M22001-32]

WARNING This product is not for human or veterinary use.