AR Zebrafish Agonist v72

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Zebrafish AR 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
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Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Zebrafish Androgen Receptor. INDIGO’s zAR reporter assay utilizes proprietary non-human cells that have been engineered to provide constitutive expression of the zAR. In addition to zAR 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 zAR. 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
SpeciesZebrafish
Receptor FormNative
Assay ModeAgonist, Antagonist
Kit Components
  • Zebrafish AR Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • 11-Ketotestosterone (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 Zebrafish AR. Agonist assays: zfAR Reporter Cells were treated with the reference agonists 11-KetoTestosterone (provided), DihydroTestosterone (DHT) and 6a-FluoroTestosterone. Luminescence/well was quantified and values of average relative light units (RLU), standard deviation (SD), percent coefficient of variation (%CV) and fold-activation were determined for each treatment concentration. Values of Fold-activation (agonist assays) or average RLU (antagonist assays) +/- %CV were plotted against their corresponding drug treatment concentrations, Log10 (pM), using GraphPad Prism software. Z’ values were calculated as described by Zhang, et al. (1999)
Antagonist dose-responses of Zebrafish AR. Antagonist assays: zfAR Reporter Cells were co-treated with a fixed EC80 concentration of the reference agonist 11-Ketotestosterone and increasing concentrations of the antagonists Hydroxy flutamide, Mifepristone, Vinclozolin, Nilutamide and bis-Phenol A (n = 3). Luminescence/well was quantified and values of average relative light units (RLU), standard deviation (SD), percent coefficient of variation (%CV) and fold-activation were determined for each treatment concentration. Values of Fold-activation (agonist assays) or average RLU (antagonist assays) +/- %CV were plotted against their corresponding drug treatment concentrations, Log10 (pM), using GraphPad Prism software. Z’ values were calculated as described by Zhang, et al. (1999)

Target Background

INDIGO’s Zebrafish Androgen Receptor Reporter Assay System utilizes proprietary mammalian cells engineered to provide constitutive, high-level expression of Zebrafish (Danio rerio) Androgen Receptor (nr3c4), a ligand-dependent transcription factor.

INDIGO’s Reporter Cells include the luciferase reporter gene functionally linked to an AR-responsive promoter. Thus, quantifying changes in luciferase expression in the test sample-treated reporter cells provides a sensitive surrogate measure of changes in zAR activity. The principal application of this assay is in the screening of test samples to quantify any functional bioactivity that they may exert against zebrafish AR. In particular, zebrafish reporter assays are frequently used in the monitoring of environmental samples for the presence of biohazardous chemical pollutants, such as endocrine disruptors.

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Androgen Receptor (AR, NR3C4)

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