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.
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.
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.
Glucocorticoid Receptor Reporter Cells
Glucocorticoid Receptor 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.
Glucocorticoid Receptor Luciferase Reporter
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.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
GR assay kit also available in: Human, Mouse, Rat, Zebrafish
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.
Service Assays: Human, Mouse, Rat, Zebrafish
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.
Glucocorticoid Receptor Assay Research Areas
Diabetes; Obesity; Inflammation; Lipid Metabolism and Energy; Metabolic Disease; Steroid Receptor;