Glucocorticoid Receptor (GR; NR3C1)
|Product Family||Product Number||Product Description||Technical Manual|
|IB00201-32||Human GR Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB00201||Human GR Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB00202||Human GR Reporter Assay System, 1 x 384-well format assays||Technical Manual|
Glucocorticoid Receptor Assay Kit
This Glucocorticoid Receptor (GR) assay kit is an all-inclusive GR reporter assay system that includes, in addition to GR 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.
GR 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 Glucocorticoid Receptor 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.
Glucocorticoid Receptor Assay Kits are offered in different assay formats to accommodate researchers' needs: 3x 32, 1x 96, and 1x 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.
Glucocorticoid Receptor 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.
Glucocorticoid Receptor Background
The glucocorticoid receptor is the receptor that cortisol and other glucocorticoids bind to. The GR is expressed in almost every cell in the body and regulates genes controlling the development, metabolism, and immune response.
INDIGO's Glucocorticoid Receptor Assay System utilize proprietary mammalian reporter cells engineered to provide constitutive high-level expression of full-length, unmodified human NR3C1 protein, commonly referred to as GR.
INDIGO's Reporter Cells include the luciferase reporter gene functionally linked to a GR-responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in GR activity. Luciferase gene expression occurs after ligand-bound GR undergoes nuclear translocation, DNA binding, recruitment, and assembly of the co-activators and accessory factors required to forma 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 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 androgen receptor.
Synonyms: Glucocorticoid Receptor, GR, Human Glucocorticoid Receptor, Human GR, NR3C1
Glucocorticoid Receptor Assay Data
Activity dose-response of GR using various reference agonists. Dose-response analyses of GR were performed according to the protocol provided in the assay Technical Manual. Reporter Cells were treated with the reference agonists Dexamethasone (provided), and Mometasone furoate, Fluticasone proprionate, Budesonide, Methylprednisolone, GSK 9027, and Hydrocortisone (all from Tocris). Luminescence was quantified using a plate-reading luminometer. 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: The EC50 for dexamethasone is approximately 260 pM. The 3 nM treatment concentration of dexamethasone yielded an 83-fold activation over the untreated reporter cells, with a corresponding Z’= 0.78. These data confirm the robust performance of this Human GR assay and demonstrate its suitability for use in HTS applications.