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Human AP-1 Reporter Assay Kit

1 x-96 well format assays
3 x-32 assays in-96 well format
1 x-96 well format assays
3 x-32 assays in-96 well format

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.


  • Clear, Reproducible Results

  • All-Inclusive Assay Systems
  • Exceptional Cell Viability Post-Thaw
  • Consistent Results Lot to Lot

Product Specifications

Target TypeTranscription Factor
Receptor FormNative
Assay ModeAgonist, Antagonist
Kit Components
  • AP-1 Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • PMA, (ref. activation; in DMSO)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Orthologs AvailableNo
Shipping RequirementsDry Ice
Storage temperature-80C


Dose-response activation of AP-1 using PMA. Activation of AP-1 is demonstrated by treating reporter cells with Phorbol 12-myristate 13-acetate (PMA; provided) for 22 hours, following the protocol for activation assays depicted in the assay technical manual. Average relative light units (RLU) and corresponding standard deviation (SD) values were determined for each treatment concentration (n = 4). Fold-activation and Z’ values were calculated as described by Zhang, et al. (1999). Non-linear regression and EC50 analyses were performed using GraphPad Prism software. High Z' scores confirm the robust performance of this assay.
Dose-dependent inhibition of AP-1. Human AP-1 Reporter Cells were treated with an ~EC80 concentration of PMA (10 nM) and then challenged with the inhibitors JNK Inhibitor V, CC-401, PD98059, CAY 10561, and PKC inhibitors Bisindolylmaleimide I and XI (all from Cayman Chemical, USA). Reporter cells were treated for 6 hours, following the protocol for inhibition assays depicted in the assay technical manual.

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.

Also available as a service

Activator Protein-1 (AP-1)

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