Estrogen Receptor Beta (ERβ; NR3A2)
|Product Family||Product Number||Product Description||Technical Manual|
|IB00411-32||Human ERβ Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB00411||Human ERβ Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB00412||Human ERβ Reporter Assay System, 1 x 384-well format assays|
Estrogen Receptor Beta Assay Kit
This Estrogen Receptor Beta (ERβ) assay kit is an all-inclusive ER beta reporter assay system that includes, in addition to ERβ 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.
ERβ 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 ERβ 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: 3 x 32, 1 x 96, and 1 x 384 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.
Estrogen Receptor Beta 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.
Estrogen Receptor Beta Background
Estrogen receptor beta (ER-beta), also known as NR3A2 (nuclear receptor subfamily 3, group A, member 2), is a nuclear receptor which is activated by the sex hormone estrogen. ER beta is encoded by the human gene ESR2 (Estrogen Receptor 2). This gene encodes a member of the family of estrogen receptors and superfamily of nuclear receptor transcription factors. The gene product contains an N-terminal DNA binding domain and C-terminal ligand binding domain and is localized to the nucleus, cytoplasm, and mitochondria. Upon binding to 17beta-estradiol or related ligands, the encoded protein forms homo- or hetero-dimers that interact with specific DNA sequences to activate transcription. Some isoforms dominantly inhibit the activity of other estrogen receptor family members. Several alternatively spliced transcript variants of this gene have been described, but the full-length nature of some of these variants has not been fully characterized. ERβ is expressed by many tissues including blood monocytes and tissue macrophages, colonic and pulmonary epithelial cells and in prostatic epithelium and in malignant counterparts of these tissues. ERβ may have anti-proliferative effects and therefore oppose the actions of ERα in reproductive tissue. ERβ may also have an important role in adaptive function of the lung during pregnancy.
INDIGO's Estrogen Receptor Beta Reporter Assay Systems utilize non-human mammalian cells engineered to express human ESR2, commonly referred to as NR3A2 or ERβ.
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 beta.
Synonyms:Estrogen Receptor Beta, ERβ, ER beta, ESR2, Estrogen Receptor 2, ERb, ESRB, ER-beta, Human Estrogen Receptor Beta, Human ER beta, NR3A2
Estrogen Receptor Beta Assay Data
Dose-response analyses of ERβ were performed according to the protocol provided in the assay Technical Manual. Reporter Cells were treated with 17β-Estradiol (provided), DPN, Linquiritigenin, WAY 200070, and FERb 033 (all from Tocris). Average relative light units (RLU) and corresponding standard deviation (SD) values were determined for each treatment concentration (n = 4). Values of Fold-Activation (Signal / Background) and Z’ were calculated as described by Zhang, et al. (1999). Non-linear regression and EC50 analyses were performed using GraphPad Prism software. RESULTS: EC50 Z’ values for 17β-estradiol confirm the robust performance of this ERβ Assay and demonstrate its suitability for use in HTS applications.
ERβ antagonist assays were performed using Tamoxifen citrate, PHTPP and (R,R)-THC (all from Tocris). ERβ Assay setup and quantification of antagonist activities were performed following as described in the assay Technical Manual. The bulk suspension of reporter cells was supplemented with a 2x-EC75 concentration of 17β-Estradiol and dispensed into the assay plate at 100 ul/well. Cells were then further treated by adding 100 ul/well of decreasing 2x-concentrations of antagonist. Assay plates were incubated for ~23 hrs, then processed to quantify ERβ activity for each treatment condition.