Progesterone Receptor (PGR; NR3C3)
|Product Family||Product Number||Product Description||Technical Manual|
|IB05001-32||Human PGR Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB05001||Human PGR Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB05002||Human PGR Reporter Assay System, 1 x 384-well format assays||Technical Manual|
Progesterone Receptor Assay Kit
This Progesterone Receptor (PGR) assay kit is an all-inclusive PGR reporter assay system that includes, in addition to PGR 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.
Progesterone Receptor Reporter Cells
Progesterone 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.
Progesterone Receptor Luciferase Reporter
INDIGO’s Progesterone 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.
PGR assay kits also available in: Human, Rat
Progesterone Receptor Assay Services
The primary application of INDIGO’s cell-based Progesterone Receptor assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of the Progesterone 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.
PGR Service Assays: Human, Rat
Progesterone Receptor Background
INDIGO’s Human Progesterone Receptor (PGR) Reporter Assay System utilizes proprietary mammalian cells engineered to provide constitutive, high-level expression of human PGR (NR3C3), a ligand-dependent transcription factor.
INDIGO’s Reporter Cells include the luciferase reporter gene functionally linked to a PGR-responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in PGR activity. Luciferase gene expression occurs after ligand-bound PGR 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 in vitro binding assays, and 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.
For more information on PGR, visit the Nuclear Receptor Resource.
Synonyms: Progesterone Receptor, PGR, Human Progesterone Receptor, Human PGR, Human PR, NR3C3
Progesterone Receptor Assay Data
Agonist dose-response analyses of Human PGR. Agonist dose-response of Human PGR Reporter Cells using Progesterone (provided), and Nomegestrol acetate (Tocris). Concentrated stocks prepared in DMSO were serially diluted in 5-fold decrements using CSM. Final assay concentrations for progesterone-treated cells ranged between 1,000 nM and 64 pM; assay concentrations of nomegestrol ranged between 40 nM and 2.6 pM. Luminescence was quantified and average relative light units (RLU) and their corresponding values of standard deviation (SD), Fold-Activation and Percent Relative Activation were determined for each treatment group (n = 3). Z’ values were calculated as described by Zhang, et al. (1999). GraphPad Prism software was used to curve-fit data using the least-squares method of non-linear regression, and EC50 values were determined. High Z' scores confirm the robust performance of this PGR Assay.
Validation of PGR Assay antagonist dose-response. Antagonist analysis of PGR Reporter Cells using the reference compounds Mifepristone (Cayman Chem.) and PF-02413873 (Sigma-Aldrich). Reporter cells were exposed to a fixed ~ EC80 concentration of the challenge agonist Progesterone and varying concentrations of the reference antagonists. All treatments were performed in triplicate. The Live Cell Multiplex (LCM) assay confirmed that no compound-induced cytotoxicity occurred (data not shown).
Progesterone Receptor Assay Services
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