Provider: INDIGO Biosciences, Inc. provides Nuclear Receptor Reporter Assay based products and services for drug discovery and toxicology. INDIGO specializes in the development of frozen, whole-cell Nuclear Receptor Assay Systems for use in small compound HTS, off-target drug interactions, and toxicology screens.
Background: The calcitriol receptor, also known as the vitamin D receptor (VDR) and also known as NR1I1 (nuclear receptor subfamily 1, group I, member 1), is a member of the nuclear receptor family of transcription factors. Upon activation by vitamin D, the VDR forms a heterodimer with the retinoid-X receptor and binds to hormone response elements on DNA resulting in expression or transrepression of specific geneproducts. In humans, the vitamin D receptor is encoded by the VDR gene. Glucocorticoids are known to decrease expression of VDR which is expressed in most tissues of the body and regulate intestinal transport of calcium. This gene encodes the nuclear hormone receptor for vitamin D3. This receptor also functions as a receptor for the secondary bile acid lithocholic acid. The receptor belongs to the family of trans-acting transcriptional regulatory factors and shows similarity of sequence to the steroid and thyroid hormone receptors. Downstream targets of this nuclear hormone receptor are principally involved in mineral metabolism though the receptor regulates a variety of other metabolic pathways, such as those involved in the immune response and cancer. Mutations in this gene are associated with type II vitamin D-resistant rickets. A single nucleotide polymorphism in the initiation codon results in an alternate translation start site three codons downstream. Alternative splicing results in multiple transcript variants encoding the same protein.
Service Details: This assay can be utilized to examine the agonism (activation) or antagonism (inhibition of an agonist) on the nuclear receptor. Mammalian cells are transfected with the ligand binding domain of the NR fused to a heterologous DNA binding domain as well as the response element for this DNA binding motif linked to a reporter gene. A transfection efficiency control plasmid is also included. These transfected cells are seeded in 96-well plates and then treated overnight with test compound at 37°C. Cells are then lysed and NR activation reporter activity as well as transfection efficiency reporter activity is assessed. A positive control compound (6 concentrations in triplicate) as well as vehicle control (triplicate) is included in every experiment.
For dose response curves, it is suggested that at least six concentrations of the test compounds be examined, covering a wide range of dilutions (3 log-units). In addition, we recommend that each compound/concentration be assayed in triplicate.
Deliverable: Downloadable data report containing raw and analyzed data. EC50 and peak activity will be determined using nonlinear regression. Dose-response data will be plotted for the test compounds as well as the positive control. A sample data report is available.
Price: The price of the assay includes a set-up fee, as well as a per well charge. Discounts are provided for academicians and for larger projects. Please contact us by email or phone (814) 234-1919 Ext. 303 for a price quote.
Sample Submission: The minimum sample requirement is 25 µl of a 10 mM stock solution, with the preferred sample submission of 100 mM stock solution in DMSO. A sample submission sheet is available.
Species available: Reporter assays for human are optimized and ready for use. Assays for other species can be custom prepared. Please visit the Custom Assay Development page for more details
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