NFAT Reporter Assay Kit
Product Description and Product Data
This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Nuclear Factor of Activated T cells (NFAT). INDIGO’s NFAT reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of NFAT. In addition to NFAT 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 NFAT. 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 | Transcription Factor | ||
| Species | Human | ||
| Receptor Form | Native | ||
| 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
The five members of the Nuclear Factor of Activated T cells (NFAT1-5) family were initially identified as key regulators of genes involved in the activation, proliferation, differentiation, and apoptosis of cells, most notably the T cells and B cells of the immune system.
Inactive NFAT resides in the cytoplasm in a multi-phosphorylated form. Phospho-NFAT is converted to its active form through the activation of calcineurin, a calcium-dependent phosphatase. Any physiological event that drives the influx of extra-cellular Ca+2, or depletes internal Ca+2 stores within the endoplasmic reticula, results in the Ca+2-activation of calcineurin and its subsequent dephosphorylation of phospho-NFAT. Activated NFAT translocates to the nucleus where it binds to specific genetic response element (GRE) sequences within the promoter / enhancer region of target genes. Functional transcription complexes form via cooperative association with other transcription factors, most notably AP-1.
Importantly, dysregulation of the calcineurin-NFAT pathway is strongly associated with autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis, thereby making it an important therapeutic target for small molecule drug development.
INDIGO’s Reporter Cells contain an engineered luciferase reporter gene functionally linked to tandem NFAT/AP-1 GRE sequences positioned immediately upstream of a minimal promoter. Activated NFAT will bind to its corresponding GREs to initiate the formation of a complete transcription complex that drives Luc gene expression. Thus, quantifying changes in luciferase activity in the treated reporter cells provides a sensitive surrogate measure of changes in NFAT activity. Accordingly, the principal application of this reporter assay is in the screening of test compounds to quantify any functional activities, either activating or inhibitory, that they may exert against the calcineurin-NFAT signal transduction pathway.
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