Zebrafish Androgen Receptor (zAR; nr3c4)
|Product Family||Product Number||Product Description||Technical Manual|
|Z03001-32||Zebrafish AR Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|Z03001||Zebrafish AR Reporter Assay System, 1 x 96-well format assays||Technical Manual|
Zebrafish Androgen Receptor Assay Kit
This Zebrafish Androgen Receptor Reporter (zAR) assay kit is an all-inclusive firefly luciferase reporter assay system that includes in addition to zAR 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.
zAR Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This cryo-preservation method yields high cell viability post-thaw, and provides the convenience of immediately dispensing healthy, division-competent reporter cells into assay plates. There is no need for intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.
INDIGO’s 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.
Kits are offered in different assay formats to accommodate researchers’ needs: 3x 32 and 1x 96 assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
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.
Zebrafish Androgen Receptor Assay Background
INDIGO’s Zebrafish Androgen Receptor Reporter Assay System utilizes proprietary mammalian cells engineered to provide constitutive, high-level expression of Zebrafish (Danio rerio) Androgen Receptor (nr3c4), a ligand-dependent transcription factor.
INDIGO's Reporter Cells include the luciferase reporter gene functionally linked to an AR-responsive promoter. Thus, quantifying changes in luciferase expression in the test sample-treated reporter cells provides a sensitive surrogate measure of changes in zAR activity. The principal application of this assay is in the screening of test samples to quantify any functional bioactivity that they may exert against zebrafish AR. In particular, zebrafish reporter assays are frequently used in the monitoring of environmental samples for the presence of biohazardous chemical pollutants, such as endocrine disruptors.
For more information on AR, visit the Nuclear Receptor Resource.
Zebrafish Androgen Receptor Assay Data
Agonist and Antagonist dose-responses of Zebrafish AR. Agonist assays: zAR Reporter Cells were treated with the reference agonists 11-KetoTestosterone (provided), DihydroTestosterone (DHT) and 6α-FluoroTestosterone. Antagonist assays: zAR Reporter Cells were co-treated with a fixed EC80 concentration of the reference agonist 11-Ketotestosterone and increasing concentrations of the antagonists Hydroxy flutamide, Mifepristone, Vinclozolin, Nilutamide and bis-Phenol A (n = 3). Luminescence/well was quantified and values of average relative light units (RLU), standard deviation (SD), percent coefficient of variation (%CV) and fold-activation were determined for each treatment concentration. Values of Fold-activation (agonist assays) or average RLU (antagonist assays) +/- %CV were plotted against their corresponding drug treatment concentrations, Log10 (pM), using GraphPad Prism software. Z’ values were calculated as described by Zhang, et al. (1999).