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Rat FXR Reporter Assay Kit

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

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Rat Farnesoid X Receptor. INDIGO’s rFXR reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the rFXR. In addition to rFXR Reporter 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 rFXR. 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

  • Clear, Reproducible Results

  • All-Inclusive Assay Systems
  • Exceptional Cell Viability Post-Thaw
  • Consistent Results Lot to Lot

Product Specifications

Target TypeNuclear Hormone Receptor, Nuclear Receptor Orthologs
SpeciesRat
Receptor FormHybrid
Assay ModeAgonist, Antagonist
Kit Components
  • Rat FXR Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • GW4064, (ref. agonist; in DMSO)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Agonist dose responses of the rat FXR Assay. Dose-response assays were performed as described in the Technical Manual. FXR reference agonists GW4064 (provided), Fexaramine and CDCA (Cayman Chemical), Obeticholic acid (OCA) and WAY-362450 (Selleckchem) were analyzed. Luminescence was quantified using a GloMax-Multi+ luminometer (Promega). Average relative light units (RLU) and corresponding standard deviation (SD) values were determined for each treatment concentration. Z’ values were calculated as described by Zhang, et al. (1999). RESULTS: The large assay window and high Z' values confirm the robust performance for this rat FXR assay.

Target Background

The Farnesoid X Receptor (FXR), also known as NR1H4 is a nuclear hormone receptor with activity similar to that seen in other steroid receptors such as estrogen or progesterone receptor, but more similar in form to PPAR, LXR, and RXR. FXR is expressed at high levels in the liver and intestine. Chenodeoxycholic acid and other bile acids are natural ligands for FXR.

Like other steroid receptors, when activated, FXR translocates to the cell nucleus, forms a heterodimer with RXR and binds to hormone response elements on DNA (FXEs) to elicit expression or transrepression of gene products. One of the primary functions of FXR activation is the suppression of cholesterol 7 alpha-hydroxylase (CYP7A1), the rate-limiting enzyme in bile acid synthesis from cholesterol. FXR does not directly bind to the CYP7A1 promoter. Rather, FXR induces expression of small heterodimer partner (SHP) which then functions to inhibit transcription of the CYP7A1 gene. In this way a negative feedback pathway is established in which synthesis of bile acids is inhibited when cellular levels are already high.

These Rat FXR Reporter Assay Systems utilize non-human mammalian cells engineered to express Mouse NR1H4 protein, commonly referred to as mFXR.

The principle application of this assay product is in the screening of test samples to quantify functional activities, either agonist or antagonist, that they may exert against the rat farnesoid x receptor.

Also available as a service

Farnesoid X Receptor (FXR, NR1H4)

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