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Estrogen Receptor Alpha (ERα; NR3A1)

Product Family Product Number Product Description Technical Manual
IB0040
ERα
(NR3A1)
IB00401-32 Human ERα Reporter Assay System, 3 x 32 assays in 96-well format Technical Manual
IB00401 Human ERα Reporter Assay System, 1 x 96-well format assays Technical Manual
IB00402 Human ERα Reporter Assay System, 1 x 384-well format assays Technical Manual

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  • Kits
  • Services
  • Background
  • Data
  • Research Areas

Estrogen Receptor Alpha Assay Kit INDIGO Assay Kit

This Estrogen Receptor Alpha (ERα) assay kit is an all-inclusive ER alpha reporter assay system that includes, in addition to ER Alpha 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.

INDIGO’s cell-based reporter assays allow scientists to detect any biological activity that their test samples may exert against a specific receptor present in the cell. They utilize firefly luciferase reporter gene technology which provides superior precision and sensitivity. Since the receptor binding controls the expression of the luciferase reporter gene, luciferase activity in the cells can be correlated directly with the activity of the receptor. The strength of an interaction of a chemical with the target receptor is quantified using a luminometer to measure the level of light emitted.

Thaw, Feed, Dose, Read

Fast, reproducible, easy-to-analyze results are only four steps away

Many luciferase reporter assays require the user to grow their own cells and take time to optimize the results. INDIGO’s reporter cells contain the receptor of interest and the luciferase reporter gene. Reporter cells have been optimized to provide extreme sensitivity to quantify even small changes in receptor activity. With INDIGO’s cell-based reporter assays, the process is as easy as Thaw, Feed, Dose, and Read.

Estrogen Receptor Alpha Reporter Cells

Estrogen Receptor Alpha Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This proprietary cryopreservation process, enables long-term preservation of our unique reporter cells, so we can ship our cryopreserved reporter cells and assay reagents to you via overnight delivery, for your immediate use. Or, you can store the assay kits at -80°C. Once thawed, reporter cells are ready for immediate use so there is no need to take time on intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.

Estrogen Receptor Alpha Luciferase Reporter

INDIGO’s Estrogen Receptor Alpha 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.

Assay Kit & Platforms

Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.

ERa assay kits also available in: Zebrafish

Estrogen Receptor Alpha Assay Services

The primary application of INDIGO’s cell-based Estrogen Receptor Alpha assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of Estrogen Receptor Alpha. 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.

Service Assays: Human, Zebrafish

Estrogen Receptor Alpha Background

ER-alpha is encoded by the human gene ESR1 (Estrogen Receptor 1). The estrogen receptor (ESR) is a ligand-activated transcription factor composed of several domains important for hormone binding, DNA binding, and activation of transcription. Alternative splicing results in several ESR1 mRNA transcripts, which differ primarily in their 5-prime untranslated regions. The translated receptors show less variability.

INDIGO's Estrogen Receptor Alpha Reporter Assay Systems utilize proprietary mammalian cells engineered to express constitutive, high-level expression of the full length Human Estrogen Receptor 1 (NR3A1), a ligand-dependent transcription factor commonly referred to as ERα.

INDIGO's Estrogen Receptor Alpha Reporter Cells include the luciferase reporter gene functionally linked to an ERα-responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in ERα activity. Luciferase gene expression occurs after ligand-bound ERα 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 target 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 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 human estrogen receptor alpha.

For more information on ERα, visit the Nuclear Receptor Resource.

Synonyms: Estrogen Receptor Alpha, ERα, ER alpha, ESR1, Estrogen Receptor 1, ERa, ESRA, ER-alpha, NR3A1

Estrogen Receptor Alpha Assay Data

Estrogen Receptor Alpha Agonist

Analyses of Human ERα Reporter Cells using 17-β-Estradiol (provided), PPT (4,4'4"-(4-Propyl-[1H]-pyrazole-1,3,5-triyl tris-phenol; Tocris), and (R,R)-THC ([R,R]-5,11-diethyl-5,6,11,12-tetrahydro-2,8-chrysenediol; Tocris). Concentrated stocks of agonists were prepared in DMSO; the highest concentration of DMSO in assay wells was 0.1%. Luminescence was quantified and values of average relative light units (RLU) and corresponding standard deviation (SD) were determined for each treatment concentration (n ≥ 3). Fold-activation and Z’ values were calculated as described by Zhang, et al. (1999). Non-linear regression curve-fitting of the data and EC50 determinations were performed using GraphPad Prism software.

Estrogen Receptor Alpha Antagonist

Human ERα antagonist assays were performed using Endoxifen, Tamoxifen citrate, ICI182780 and MPP dihydrochloride (all from Tocris). INDIGO’s Live Cell Multiplex Assay (#LCM-01) was also performed to confirm that the observed drop in RLU values resulted from receptor-specific inhibition, and not from compound-induced cell death (data not shown). Reporter cells were co-treated with a fixed concentration (3.2 nM, approximating EC75) of the agonist 17β-Estradiol and varying concentrations of respective challenge antagonists.

Estrogen Receptor Alpha Assay Research Areas

Steroid Receptor; Osteoporosis; Cancer; Fertility; Alzheimer’s Disease; Cardiovascular Disease; Reproduction and Development; Endocrinology; Autoimmune Disease; Environmental Toxicology

Research articles associated with ERα

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