AR Reporter Assay System (dog)

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Dog Androgen Receptor (dAR). INDIGO's dAR reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the dAR. In addition to dAR 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 dog AR. 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 Ready to Use Upon Receipt Includes All Needed Components Contains Transfected Reporter Cells Eliminates Cell Licensing Fees Clear, Reproducible Results Consistent Results Lot to Lot Product Specifications Target Type Nuclear Hormone Receptor, Nuclear Receptor Orthologs Species Dog Receptor Form Hybrid Assay Mode Agonist, Antagonist Kit Components Dog 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 Life 6 months Orthologs Available Yes. Target Background: This AR assay utilizes proprietary cells engineered to provide constitutive, high-level expression of full length, unmodified Dog 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. Thus, 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 principal 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 dog androgen receptor. It is an all-inclusive assay system that includes, in addition to AR Reporter Cells, two optimized media for use during cell culture and for preparing dilutions of test samples, the reference agonist 5alphaDihydro 11-keto Testosterone, Luciferase Detection Reagent, a cell culture-ready assay plate, and a detailed protocol. Protein function: Steroid hormone receptors are ligand-activated transcription factors that regulate eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Transcription factor activity is modulated by bound coactivator and corepressor proteins like ZBTB7A that recruits NCOR1 and NCOR2 to the androgen response elements/ARE on target genes, negatively regulating androgen receptor signaling and androgen-induced cell proliferation. Transcription activation is also down-regulated by NR0B2. Activated, but not phosphorylated, by HIPK3 and ZIPK/DAPK3. [The UniProt Consortium]