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Rat AR 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 Androgen Receptor. INDIGO’s rAR reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the rAR. In addition to rAR 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 rAR. 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 AR Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • 5a-Dihydro-11-keto Testosterone; (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. Rat AR agonist dose-response assays were performed using the reference agonists 5aDihydro 11-keto Testosterone (5aDH-11kT; provided), Dihydro Testosterone (DHT; Steraloids Inc.), 6a-Fl Testosterone (6aFlT; Enzo Life Sci), Cl-4As-1, 11-keto Testosterone (11-kT; Sigma-Aldrich) and Medroxy-Progesterone 17-Acetate (MPA; Steraloids, Inc.). Luminescence was quantified and values of average relative light units (RLU), corresponding standard deviation (SD), Fold-Activation and Z’ were determined. Non-linear regression analyses of Fold-Activation vs. Log10 transformed concentration were plotted and EC50 values determined using GraphPad Prism software

Target Background

The androgen receptor is activated by binding of either of the androgenic hormones testosterone or dihydrotestosterone. The androgen receptor is most closely related to the progesterone receptor, and progestins in higher dosages can block the androgen receptor. The main function of the androgen receptor is as a DNA binding transcription factor which regulates gene expression; however, the androgen receptor has other functions as well.

Androgen regulated genes are critical for the development and maintenance of the male sexual phenotype. In some cell types testosterone interacts directly with androgen receptors while in others testosterone is converted by 5-alpha-reductase to dihydrotestosterone, an even more potent agonist for androgen receptor activation. Testosterone appears to be the primary androgen receptor activating hormone in the Wolffian duct while dihydrotestosterone is the main androgenic hormone in the urogenital sinus, urogenital tubercle, and hair follicles. Hence testosterone is primarily responsible for the development of male primary sexual characteristics while dihydrotestosterone is responsible for secondary male characteristics.

This nuclear receptor assay system utilizes proprietary non-human mammalian cells engineered to provide constitutive, high-level expression of full length, unmodified Rat Androgen Receptor (nr3c4; AR), a ligand-dependent transcription factor commonly referred to as rAR.

INDIGO’s Reporter Cells include the luciferase reporter gene functionally linked to an AR-responsive promoter. Therefore, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in rAR activity. Luciferase gene expression occurs after ligand-bound rAR 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 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 rat androgen receptor.

Also available as a service

Androgen Receptor (AR, NR3C4)

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