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Rat PGR Reporter Assay Kit

SIZE SKU PRICE
1 x-96 well format assays
3 x-32 assays in-96 well format
SIZE SKU
1 x-96 well format assays
3 x-32 assays in-96 well format

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Rat Progesterone Receptor. INDIGO’s rPGR reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the rPGR. In addition to rPGR Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist, Luciferase Detection Reagent, and a cell culture-ready assay plate. The principal application of this assay is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against rPGR. This kit provides researchers with clear, reproducible results, exceptional cell viability post-thaw, and consistent results lot to lot. Kits must be stored at -80C. Do not store in liquid nitrogen. Note: reporter cells cannot be refrozen or maintained in extended culture.

Features

  • Clear, Reproducible Results

  • All-Inclusive Assay Systems
  • Exceptional Cell Viability Post-Thaw
  • Consistent Results Lot to Lot

Product Specifications

Target TypeNuclear Hormone Receptor, Nuclear Receptor Orthologs
SpeciesRat
Receptor FormNative
Assay ModeAgonist, Antagonist
Kit Components
  • Rat PGR Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Progesterone, (ref. agonist; in DMSO)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Agonist dose-response analyses of Rat PGR. Agonist analyses of rPGR Reporter Cells using progesterone (provided), nomegestrol acetate and mometasone furoate (Tocris), and melengestrol acetate (Enzo). 1,000x-concentrated stocks were prepared in DMSO, then serially diluted in CSM using 3-fold decrements. Average relative light units (RLU) and corresponding standard deviation (SD) values were determined for each treatment concentration (n =3). Fold-activation (i.e., signal-to-background) and Z’ values were calculated as described by Zhang, et al. (1999). Non-linear regression and EC50 analyses were performed using GraphPad Prism software. The high Z' score confirms the robust performance of this PGR Assay.
Validation of PGR Assay antagonist dose-response. PGR reference antagonists Mifepristone (Tocris) and PF-02413873 were assessed. Antagonist mode assays were setup as described in this Technical Manual. CSM was first supplemented with an EC80 concentration of the challenge agonist progesterone, and this medium was then used to prepare a series of 3-fold serial dilutions of each antagonist.

Target Background

INDIGO’s Rat Progesterone Receptor (rPGR) Reporter Assay System utilizes proprietary mammalian cells engineered to provide constitutive, high-level expression of rat PGR (NR3C3), a ligand-dependent transcription factor.

INDIGO’s Reporter Cells include the luciferase reporter gene functionally linked to a rPGR-responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in rPGR 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.

Also available as a service

Progesterone Receptor (PGR, NR3C3)

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