Fibroblast Growth Factor Receptor 1c and β-Klotho (FGFR/β-Klotho)for Endocrine FGF Signaling
|Product Family||Product Number||Product Description||Technical Manual|
|IB22001-32||Human FGFR/β-Klotho Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB22001||Human FGFR/β-Klotho Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB22002||Human FGFR/β-Klotho Reporter Assay System, 1 x 384-well format assays|
Human Fibroblast Growth Factor Receptor 1c and β-Klotho Assay Kit
This Human Fibroblast Growth Factor Receptor 1c and β-Klotho (FGFR/β-Klotho) assay kit is an all-inclusive firefly luciferase reporter assay system that includes in addition to FGFR/β-Klotho Reporter Cells, 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.
FGFR/β-Klotho Reporter Cells 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 cumbersome intermediate treatment steps such as spin-and-rinse of cells, viability determinations, or cell titer adjustments prior to assay setup.
INDIGO’s FGFR/β-Klotho assay kits feature a luciferase detection reagent specially formulated to provide stable light emission between 5 and 90+ minutes after initiating the luciferase reaction. Incorporating a 5-minute reaction-rest period ensures that light emission profiles attain maximal stability, thereby allowing assay plates to be processed in batch. By doing so, the signal output from all sample wells, from one plate to the next, may be directly compared within an experimental set.
FGFR/β-Klotho assay kits are offered in different assay formats to accommodate researchers' needs: 3x 32, 1x 96, and 1x 384 assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
Human Fibroblast Growth Factor Receptor 1c and β-Klotho Assay Services
The primary application of INDIGO's cell-based FGFR21 assays are to quantitatively assess the bioactivity of a test compound as an agonist or antagonist of the receptor. Service assays include a positive control reference compound and 'vehicle' control for every experiment. A formal study report and all data files are provided to the client upon completion of the study. To receive a quote for your proposed study, complete & submit the online "Request a Quote" form or contact an INDIGO Customer Service Representative to discuss your desired study parameters.
Human Fibroblast Growth Factor Receptor 1c and β-Klotho Assay Background
The family of Fibroblast Growth Factors (FGFs) comprise approximately 23 members that are related by core sequence and structure conservation, with the majority of FGFs being secreted signaling proteins. Secreted FGFs are predominantly autocrine and paracrine factors, with only three members evolved to function as endocrine factors. FGFs bind and activate FGF Receptors (FGFRs) which, themselves, are members of the family of high-affinity tyrosine kinase receptors.
Paracrine FGFs show high affinity towards the extracellular matrix (ECM) component heparin sulfate (HS) and are thus retained in the ECM and function locally. In contrast, the atypical endocrine subfamily of FGFs, that comprise FGF-19, FGF-21, and FGF-23, have reduced affinity for HS and can therefore escape from the ECM into the circulation to reach target distant organs.
FGF-21 a liver secreted endocrine FGF, has been identified as a factor that has multiple beneficial effects on obesity-related disorders. For example, it is a potent activator of glucose uptake in primary human adipocytes. In addition, FGF-21 has been shown to protect diet-induced obesity and diabetic illness in animals. Consequently, the associated metabolic benefits of the endocrine FGFs have promoted considerable interest in therapeutic development and drug safety screening.
INDIGO's Human Fibroblast Growth Factor Receptor Receptor 1c and β-Klotho (FGFR/β-Klotho) assay utilizes proprietary human cells that have been engineered to provide constitutive expression of the Human Fibroblast Growth Factor Receptor 1c and β-Klotho. FGFR1c and β-Klotho are both single-pass transmembrane proteins. FGFR1c has an extracellular ligand-binding domain, transmembrane domain, and intracellular tyrosine kinase domain. It has been established that FGFR 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. It is FGFR1c/β-Klotho signal transduction via the RAS-MAPK-ERK1/2 cascade that is exploited by the reporter cells in this assay.
INDIGO's FGFR/β-Klotho Reporter Cells contain the luciferase reporter gene functionally linked to an Elk-1 responsive promoter. Activated Elk-1 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 FGFR1c/β-Klotho activity.
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 FGFR1c/β-Klotho or the coupled RAS-MAPK pathway.
Synonyms: Fibroblast Growth Factor Receptor, Fibroblast Growth Factor 21, FGF 21, Fibroblast Growth Factor Receptor 1c and β-Klotho, FGFR1c/β-Klotho, FGFR1c
Human Fibroblast Growth Factor Receptor 1c and β-Klotho Assay Research Areas
NAFLD/NASH; Inflammation; Diabetes; Obesity; Cardiovascular; Metabolic Disease