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Androgen Receptor (AR; NR3C4)

Product Family Product Number Product Description Technical Manual
IB0300
AR
(NR3C4)
IB03001-32 Human AR Reporter Assay System, 3 x 32 assays in 96-well format Technical Manual
IB03001 Human AR Reporter Assay System, 1 x 96-well format assays Technical Manual
IB03002 Human AR Reporter Assay System, 1 x 384-well format assays Technical Manual

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  • Kits
  • Services
  • Background
  • Data
  • Research Areas

Androgen Receptor Assay Kits INDIGO Assay Kit

This Androgen Receptor (AR) assay kit is an all-inclusive AR reporter assay system that includes, in addition to AR 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.

Androgen Receptor Reporter Cells

Androgen 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.

Androgen Receptor Luciferase Reporter

INDIGO’s Androgen 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.

Assay kits also available in: Rat, Zebrafish

Androgen Receptor Assay Services

The primary application of INDIGO's cell-based nuclear receptor assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of a given 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.

Androgen Service Assays: Human, Rat, Zebrafish

Androgen Receptor 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.

INDIGO's Androgen Receptor assay system utilizes proprietary mammalian cells engineered to provide constitutive, high-level expression of full length, unmodified Human Androgen Receptor (NR3C4), a ligand-dependent transcription factor commonly referred to as AR.

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

Synonyms: Androgen Receptor, AR, Dihydrotestosterone Receptor. Human Androgen Receptor, hAR, NR3C4

Androgen Receptor Assay Data

Androgen receptor agonist  dose response Data Graphs

Agonist dose-response analyses. Human AR agonist dose-response assays were performed using the reference agonists 5αDihydro 11-keto Testosterone (5αDH-11kT; provided), Dihydro Testosterone (DHT; Steraloids Inc.), 6α-Fl Testosterone (6αFlT; 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

Androgen receptor antagonist dose response Data Graph

Analyses of AR Reporter Cells treated with 100 pM 5αDihydro-11-keto Testosterone (~EC70) and challenged with the AR reference antagonists Mifepristone (Enzo Biochem) Nilutamide or Hydroxy-Flutamide (Sigma). No compound-induced cytotoxicity was detected.

Androgen Receptor Assay Research Areas

Endocrinology; Cancer; Steroid Receptor; Osteoporosis; Reproduction and Development; Environmental Toxicology

Research articles associated with AR

 

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