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Rat ERa 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 Estrogen Receptor Alpha (ER). INDIGO’s rER Alpha reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the rER Alpha. In addition to rER 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 rER 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.

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 FormNative
Assay ModeAgonist, Antagonist
Kit Components
  • Rat 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

Data

Agonist dose-response analyses of the Rat ERa assay. Agonist-mode dose-response analysis of the rat ERa were performed using the reference agonist 17b-Estradiol (E2; provided) as described in this Technical Manual. Relative Light Units (RLU) were quantified and average values of RLU, standard deviation (SD) and Fold-Activation were determined for each treatment concentration (n=3). Non-linear regression analyses of Fold-Activation vs Log10[E2, pM] and EC50 determination was performed using GraphPad Prism software. The high Z' score confirms the robust performance of this rat ERa Assay.
Antagonist dose-response analyses of the Rat ERα assay. Antagonist-mode dose-response analyses of the rat ERα were performed by co-treating the reporter cells with a fixed EC80 concentration of the agonist 17β-Estradiol (provided) and varying concentrations of the respective reference antagonists MPP dihydrochloride (Tocris), Endoxifen, Tamoxifen citrate, PHTPP, or ICI 182,780 (all from Cayman Chemical).

Target Background

INDIGO’s Rat Estrogen Receptor Alpha Reporter Assay System utilizes proprietary non-human cells engineered to provide constitutive, high-level expression of the full-length Rattus norvegicus Estrogen Receptor 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 rERα activity. Luciferase gene expression occurs after ligand-bound rERα 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 rat ERa.

Also available as a service

Estrogen Receptor Alpha (ERa, NR3A1)

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