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Progesterone Receptor (PGR; NR3C3)

Product Family Product Number Product Description Technical Manual
IB0500
PGR
(NR3C3)
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

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  • Kits
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  • Background
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Progesterone Receptor Assay Kit INDIGO 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.

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.

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.

Assay Kit & Platforms

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

Progesterone Receptor agonist assay

Agonist dose-response analyses of Human PGR. Agonist analyses of PGR Reporter Cells using Progesterone (provided), and Nomegestrol acetate (Tocris). In addition, to assess the level of background signal contributed by non-specific factors that may cause activation of the luciferase reporter gene, “mock” reporter cells were treated with Progesterone (mock reporter cells, which contain only the luciferase vector, are not provided with assay kits). 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 1,000 nM and 2.4 pM. Luminescence was quantified and average relative light units (RLU) and corresponding standard deviation (SD) and Fold-Activation values were determined for each treatment concentration (n ≥ 6). Z’ values were calculated as described by Zhang, et al. (1999). Non-linear regression and EC50 analyses were performed using GraphPad Prism software. Mock reporter cells demonstrate no significant background luminescence (< 0.1% that of the reporter cells at ECMax). Thus, luminescence results strictly through ligand-activation of PGR expressed in these reporter cells. High Z' scores confirm the robust performance of this PGR Assay.

PGR antagonist assay graphic

Validation of PGR Assay antagonist dose-response. Antagonist analysis of PGR Reporter Cells using Mifepristone (Tocris). Assay setup and quantification of PGR activity were performed following Protocol Variation 2 in the assay Technical Manual. To confirm that the observed dose-dependent increase in % inhibition resulted from PGR inhibition, not induced cell death, the relative numbers of live cells in each assay well were determined at the end of the treatment period using INDIGO's Live Cell Multiplex (LCM) Assay (#LCM-01). In brief: CSM was first supplemented with a 2x-EC75 concentration of progesterone. This medium was then used to prepare a 10-point, serial 4-fold dilution series of mifepristone to generate a range of 2x-concentration treatment media. Frozen PRReporter Cells were then thawed in CRM, and 100 µl of this cell suspension was dispensed into each well of the assay plate. Next, 100 µl of the prepared series of 2x-concentration treatment media were dispensed per well, combining with the reporter cells. The final assay concentration of mifepristone ranged between 10 µM and 40 pM, including a 'no antagonist' control (n ≥ 6 per treatment; highest [DMSO] < 0.1% f.c.). Each treatment also contained an assay concentration of 8.7 nM (~ EC75) progesterone as challenge agonist. Assay plates were incubated for ~23 hrs, then processed according to the LCM Assay protocol to quantify relative numbers of live cells per treatment condition. Plates were then further processed to quantify PGR activity for each treatment condition. Results: Mifepristone produced a dose-dependent increase in % inhibition of progesterone. The LCM Assay reveals no significant variance in the numbers of live cells per assay well, even up to the maximum treatment concentration of 10 µM. Hence, the measured increase in % inhibition of PGR activity can be attributed to dose-dependent inhibition of the progesterone receptor, and not to compound-induced cell death.

Progesterone Receptor Assay Services

Steroid Receptor; Osteoporosis; Reproduction and Development; Autoimmune Disease; Endocrinology; Environmental Toxicology

Research articles associated with PGR

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