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Rat GR 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
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Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Rat Glucocorticoid Receptor (rGR). INDIGO’s rGR reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the rGR. In addition to rGR 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 rGR. 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 GR Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Dexamethasone, (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

Response of the Rat GR to reference Agonists. Dose-response analyses of rat GR Reporter Cells were performed using 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 average fold-activation and %CV of SD were plotted via non-linear regression, and EC50 determinations were performed using GraphPad Prism software. Z’ were calculated as described by Zhang, et al. (1999). RESULTS: The EC50 for dexamethasone is approximately 1.9 nM. The 50 nM treatment concentration of dexamethasone yielded a 74-fold activation of rGR over the untreated reporter cells, with a corresponding Z’= 0.86. These data confirm the robust performance of this rat GR assay.

Target 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 Rat Glucocorticoid Receptor (nr3c1) nuclear receptor assay utilizes proprietary mammalian cells engineered to provide constitutive high-level expression of full-length, unmodified Rat Glucocorticoid Receptor (nr3c1), a ligand-dependent transcription factor, commonly referred to as GR.

DNA binding, recruitment and assembly of the co-activators and accessory factors required to form a 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.

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Glucocorticoid Receptor (GR, NR3C1)

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