Mouse Constitutive Androstane Receptor (mCAR; nr1i3)
INDIGO’s Mouse Constitutive Androstane Receptor (nr1i3) Assay is a cell-based genetic reporter assay that utilizes proprietary non-human mammalian cells engineered to provide constitutive, high-level expression of a ligand-dependent transcription factor commonly referred to as mCAR. These reporter cells utilize a modified version of mCAR in which the native N-terminal DNA binding domain (DBD) has been replaced with that of the GAL4-DBD. The mCAR ligand binding domain (LBD) is unaltered and fully functional. The reporter cells also incorporate a luciferase cDNA functionally linked to the GAL4-upstream activation sequence (UAS). Thus, quantifying expressed luciferase activity provides a sensitive surrogate measure of changes in mCAR activity resulting from direct interaction between a treatment compound and the nuclear receptor.
Because this assay system expresses the [GAL4-DBD + mCAR LBD] hybrid receptor, the activity of modulators that act through indirect mechanisms (such as those that alter the phosphorylation status of the native N-terminal amino acid sequence of the CARs) may be dampened or go undetected.
Contrary to its name, mouse CAR is not constitutively active, rather, it exhibits ligand-dependent activation. The primary application of this reporter assay system is in the screening of test compounds to quantify any functional activity, either agonist or antagonist, that they may exert on mouse CAR.
For more information on CAR, visit the Nuclear Receptor Resource.
Also available in: Human,
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. Assay systems are all inclusive, providing reporter cells, optimized growth media, media for diluting test compounds, a positive-control agonist, luciferase detection reagent, a white assay plate, a detailed protocol, and a protocol quick guide. All kits are shipped on dry ice.
mCAR 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.
This kit product is an all-inclusive assay system that includes, in addition to mCAR Reporter Cells, two optimized media for use during cell culture and (optionally) in diluting the test samples, a reference agonist (TCPOBOP), Luciferase Detection Reagent, a cell culture-ready assay plate, and a detailed protocol.
|Product Family||Product Number||Product Description|
|M0090 mCAR (nr1i3)||M00901-32||Mouse CAR Reporter Assay System, 3 x 32 assays in 96-well format|
|M00901||Mouse CAR Reporter Assay System, 1 x 96-well format assays|
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. To initiate a Service Study, download and complete all fields of the Excel worksheet “Service Work Order" then submit the electronic file to INDIGO Customer Service.
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View Full Size Research conducted by: Koji Toyokawa (1), Ewa Maddox (1), Jack Vanden Huevel (1,2), & Bruce Sherf (1) (1) INDIGO Biosciences, Inc., 1981 Pine Hall Rd, State College, PA, USA (2) Center for Molecular Toxicology and Carcinogenesis, 325 Life Sciences Building, Penn State University, University Park, PA 16802, USA Date of Publication: 2017
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