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Human AP-1 Reporter Assay Kit
Product Description and Product Data
This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Activator Protein-1 (AP-1). INDIGO’s AP-1 reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the Human Activator Protein-1 (AP-1). In addition to AP-1 Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference activator, Luciferase Detection Reagent, and a cell culture-ready assay plate. 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 AP-1. 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
AP-1 is a dimeric transcription factor composed of homodimers or heterodimers of members of the JUN (c-Jun, JunB and JunD), FOS (c-Fos, FosB, Fra-1 and Fra-2), ATF (ATF2, ATF3,/LRF1, ATF4, ATF5, ATF6B, ATF7, B-ATF, B-ATF3, JDP1 and JDP2) and MAF (MafA, MAfB, cMAf, Nrl and MafF/G.K) families of proteins. These proteins share a characteristic basic leucine zipper (bZIP) DNA-binding domain. The leucine zipper drives the dimer formation, while the basic region is required for binding to the DNA response element. The variety of AP-1 dimer configurations confer versatility to this transcription factor in regulating numerous physiological (cell proliferation, differentiation, apoptosis, migration) and pathological activities (cancer, inflammation, transplant rejection) in the cell.
The most studied form of AP-1 transcription factor is c-Jun/Fos heterodimer. Upon activation, this heterodimer binds a palindromic DNA binding sequence referred to as the TPA response element (TRE).
AP-1 is activated by a variety of physiological and environmental stimuli such as growth factors, cytokines, stress, ultraviolet radiation, and bacterial and viral infections. The dimer composition of AP-1, the cell type and the stage of development are all important in the regulation of AP-1 activity. Upon stimulation, a cascade of kinases is activated leading ultimately to the activation of MAPK, ERK and/or JNK, which in turn regulate Fos and Jun on the transcriptional and post-translational level.
Reporter Cells contain an engineered luciferase reporter gene functionally linked to tandem TPA response element (TRE) sequences positioned immediately upstream of a minimal promoter. Activated AP-1 binds to its TRE to drive Luc gene expression. Thus, quantifying changes in luciferase activity in the treated reporter cells provides a sensitive surrogate measure of changes in AP-1 activity
Therefore, the principal application of this assay is in the screening of test samples to quantify any functional activities, either activating or inhibitory, that they may exert against AP-1.
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