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Human FGFR3c/β-Klotho Reporter Assay Kit

SIZE SKU PRICE
1 x-96 well format assays—
3 x-32 assays in-96 well format—
SIZE SKU
3 x-32 assays in-96 well format
1 x-96 well format assays

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Fibroblast Growth Factor Receptor 3c and β-Klotho (FGFR3c/β-Klotho). INDIGO’s FGFR3c/β-Klotho reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the Human Fibroblast Growth Factor Receptor 3c and β-Klotho. In addition to FGFR3c/β-Klotho Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist, Luciferase Detection Reagent, and a cell culture-ready assay plate. The principal application of this assay is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against human FGFR3c/β-Klotho. This kit provides researchers with clear, reproducible results, exceptional cell viability post-thaw, and consistent results lot to lot. Kits must be stored at -80C. Do not store in liquid nitrogen. Note: reporter cells cannot be refrozen or maintained in extended culture.

Features

  • Ready to Use Upon Receipt

  • Includes All Needed Components
  • Contains Transfected Reporter Cells
  • Eliminates Cell Licensing Fees
  • Clear, Reproducible Results
  • Consistent Results Lot to Lot

Product Specifications

Target TypeGrowth Factor Receptor
SpeciesHuman
Receptor FormHybrid
Assay ModeAgonist, Antagonist
Kit Components
  • FGFR3c/β-Klotho Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • FGF-21, 200ug/ml (in PBS+0.1%BSA)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Activation of FGFR3c/β-Klotho. Activation assays were performed using the endocrine reference activator FGF-21 (provided; Peprotech). For Activation assays luminescence was quantified and values of average (n=3) relative light units (RLU), corresponding standard deviation (SD), Fold-Activation, and Z’ values were calculated. GraphPad Prism software was used to plot data using the least-squares method of non-linear regression for Fold-Activation vs. Log10 [Cmpd], and to determine EC50 values.

Target 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 organs2. However, this subfamily typically requires association with members of the Klotho family of proteins as cofactors for efficient binding to their cognate receptor(s).

Recently, FGF-21, a liver secreted 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.

Citations

Also available as a service

Fibroblast Growth Factor 3c/β-Klotho

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