Rat Estrogen Receptor Alpha (rERα; nr3a1)
|Product Family||Product Number||Product Description||Technical Manual|
|R00401-32||Rat ERα Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|R00401||Rat ERα Reporter Assay System, 1 x 96-well format assays||Technical Manual|
Rat Estrogen Receptor Alpha Assay Kit
This Rat Estrogen Receptor Alpha (rERα) assay kit is an all-inclusive firefly luciferase reporter assay system that includes in addition to rERα Reporter Cells, two optimized media for use during cell culture and (optionally) in diluting the test samples, a reference agonist, Luciferase Detection Reagent, a cell culture-ready assay plate, and a detailed protocol.
rERα Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This cryo-preservation method yields high cell viability post-thaw, and provides the convenience of immediately dispensing healthy, division-competent reporter cells into assay plates. There is no need for intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.
INDIGO’s assay kits feature a luciferase detection reagent specially formulated to provide stable light emission between 5 and 90+ minutes after initiating the luciferase reaction. Incorporating a 5-minute reaction-rest period ensures that light emission profiles attain maximal stability, thereby allowing assay plates to be processed in batch. By doing so, the signal output from all sample wells, from one plate to the next, may be directly compared within an experimental set.
Kits are offered in different assay formats to accommodate researchers’ needs: 3x 32 and 1x 96 assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
ERa assay kits also available in: Human
Rat Estrogen Receptor Alpha Assay Services
The primary application of INDIGO’s cell-based nuclear receptor assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of a given receptor. Service assays include a positive control reference compound and ‘vehicle’ control for every experiment. A formal study report and all data files are provided to the client upon completion of the study. To receive a quote for your proposed study, complete & submit the online “Request a Quote” form or contact an INDIGO Customer Service Representative to discuss your desired study parameters.
Rat Estrogen Receptor Alpha Assay 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.
For more information on ERα, visit the Nuclear Receptor Resource.
Rat Estrogen Receptor Alpha Assay Data
Agonist dose-response analyses of the Rat ERα assay. Agonist-mode dose-response analysis of the rat ERα were performed using the reference agonist 17β-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 ERα 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).