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Human CAR-3 Reporter Assay Kit
Product Description and Product Data
This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Constitutive Androstane Receptor-3 (CAR-3). INDIGO’s CAR-3 reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the CAR-3. In addition to CAR-3 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 human CAR-3. 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
Clear, Reproducible Results
- All-Inclusive Assay Systems
- Exceptional Cell Viability Post-Thaw
- Consistent Results Lot to Lot
Product Specifications
| Target Type | Nuclear Hormone Receptor | ||
| Species | Human | ||
| Receptor Form | Hybrid | ||
| Assay Mode | Agonist, Antagonist | ||
| Kit Components |
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| Shelf Life | 6 months | ||
| Orthologs Available | No | ||
| Shipping Requirements | Dry Ice | ||
| Storage temperature | -80C |
Data
Target Background
INDIGO’s Human Constitutive Androstane Receptor, isoform 3 (CAR3) Reporter Assay System utilizes proprietary mammalian cells engineered to provide high-level expression of human CAR3 (NR1I3 isoform 3), a ligand-activated transcription factor. The Reporter Cells utilize a modified version of human CAR3 in which the N-terminal DNA binding domain (DBD) has been replaced with the GAL4-DBD. The human CAR3 ligand binding domain (LBD) is unaltered and fully functional. 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 CAR3 activity resulting from a direct interaction between a treatment compound and the nuclear receptor. Because this assay system expresses a GAL4-DBD + hCAR3 LBD hybrid receptor, the bio-activity of activators that act through indirect mechanisms (such as those that alter the phosphorylation status of the native N-terminal amino acid sequence of CAR3) may be dampened or go undetected.
Human CAR 3 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 human CAR3.
Unlike most nuclear receptors, this transcriptional regulator is constitutively active in the absence of ligand but is regulated by both agonists and inverse agonists. Ligand binding results in translocation of this protein to the nucleus, where it activates or represses target gene transcription. These ligands include bilirubin, a variety of foreign compounds, steroid hormones, and prescription drugs.
Citations
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