Mouse Constitutive Androstane Receptor Reporter Assay System (C08-0339-202)

This assay utilizes proprietary non-human mammalian cells that express mouse constitutive androstane receptor (nr1I3), a ligand-dependent transcription factor referred to as mCAR. These reporter cells utilize a modified version of mCAR in which the N-terminal DNA binding domain (DBD) has been replaced with 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 a 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 samples to quantify any functional activity, either agonist or antagonist, that they may exert on mouse CAR. 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 Bioscience’s nuclear receptor reporter assays are all-inclusive cell-based assay systems. In addition to mCAR reporter cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist (TCPOBOP), luciferase detection reagent, and a cell culture-ready assay plate.