AR Bioassay for Environmental Sample Monitoring

This Androgen Receptor Bioassay for Environmental Sample Monitoring kit is an all-inclusive luciferase reporter assay system that includes, in addition to AR Reporter Cells, two optimized media for use during cell culture and in diluting the user's test samples, Luciferase Detection Reagent, and 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. AR Bioassay Reporter Cells AR Bioassay Reporter Cells are prepared using INDIGO's proprietary CryoMite(TM) 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. AR Bioassay Luciferase Reporter INDIGO's AR Bioassay 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. AR Bioassay for Environmental Sample Monitoring Background: The principal application of this assay is to functionally screen processed water samples, with the aim of quantifying the bioactivities, if any, of contaminating endocrine disrupting chemical (EDC) entities. This assay utilizes proprietary mammalian cells expressing the native Androgen Receptor (AR, NR3C4), a ligand-dependent transcription factor. The cells also include the luciferase reporter gene functionally linked to an AR-responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in total AR activity. 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 (PubMed:20812024). Transcription activation is also down-regulated by NR0B2. Activated, but not phosphorylated, by HIPK3 and ZIPK/DAPK3. [The UniProt Consortium]