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Zebrafish ERa Reporter Assay Kit

1 x-96 well format assays
3 x-32 assays in-96 well format
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 Zebrafish Estrogen Receptor alpha. INDIGO’s zER alpha reporter assay utilizes proprietary non-human cells that have been engineered to provide constitutive expression of the zER alpha. In addition to zER alpha 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 zER alpha. 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.


  • 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
Receptor FormNative
Assay ModeAgonist, Antagonist
Kit Components
  • Zebrafish ERa Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • 17b-Estradiol, (ref. agonist; in DMSO)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C


Agonist and Antagonist dose-response analyses of Zebrafish ERa. zfERa Reporter Cells were treated with the agonists 17b-Estradiol (provided) and (R,R)-THC and PPT. For antagonist assays reporter cells were co-treated with a fixed (EC80) concentration of 17-estradiol and varying concentrations of the antagonists Endoxifen, ICI 182780, MPP and Tamoxifen citrate (all from Tocris). Assay workflow was as described in this Technical Manual. Luminescence/well was quantified and the average relative light units (RLU), corresponding standard deviation (SD), percent coefficient of variation (%CV) and fold-activation values were determined for each treatment concentration (n = 4). Average fold-activation +/- %CV (agonist assays) or RLU +/- SD (antagonist assays) were plotted against their respective treatment concentrations, Log10 (nM), using GraphPad Prism software

Target Background

INDIGO’s Zebrafish Estrogen Receptor Alpha Reporter Assay System utilizes proprietary non-human cells engineered to provide constitutive, high-level expression of Zebrafish (Danio rerio) Estrogen Receptor Alpha 1 (nr3a1), a ligand-dependent transcription factor.

INDIGO’s Reporter Cells include the luciferase reporter gene functionally linked to an ERα-responsive promoter. Thus, quantifying changes in luciferase expression in the test sample-treated reporter cells provides a sensitive surrogate measure of changes in zERα activity. Luciferase gene expression occurs after ligand-bound zERα undergoes nuclear translocation, DNA binding, recruitment and assembly of the co-activators, and accessory factors required to form a functional transcription complex, culminating in expression of the reporter gene. Unlike some other cell-based assay strategies, the readout from INDIGO’s reporter cells demands the same orchestration of all intracellular molecular interactions and events that can be expected to occur in vivo.

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.

Also available as a service

Estrogen Receptor Alpha (ERa, NR3A1)

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