View the slide viewer

Have questions about this assay kit?

Human TEAD4/YAP Assay Kit

1 x-96 well format assays
3 x-32 assays in-96 well format
1 x-384 well format assays
1 x-96 well format assays
3 x-32 assays in-96 well format
1 x-384 well format assays

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human TEAD4/YAP (Hippo Pathway). INDIGO’s TEAD4/YAP reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of TEAD4/YAP. In addition to TEAD4/YAP 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 TEAD4/YAP. 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 FormHybrid
Assay ModeInverse Agonist
Kit Components
  • Tead4/YAP Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Flufenamic Acid, (ref. inhibitor; in DMSO)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Orthologs AvailableNo
Shipping RequirementsDry Ice
Storage temperature-80C


The TEAD4/YAP Reporter Cell suspension was dispensed into a 96-well assay plate, which then underwent a preliminary 4-hour incubation. The pre-incubation media were discarded, and the following preparations of treatment media were added to the assay wells (n = 4 / treatment): -DMSO (‘vehicle only’ Control treatment) -Flufenamic Acid (binds to TEAD to inhibit TEAD/YAP transcription) -Verteporfin (promotes YAP degradation) -AICAR (promotes phosphorylation of YAP via AMPK3 → degradation) -XMU-MP-1 (cell-type specific interaction with YAP) -TED-347 (Binds to TEAD to inhibit TEAD/YAP transcription activity) After 22 hr incubation (44 hr incubation for TED-347) treatment media were discarded, Luciferase Detection Reagent was added, and RLU/well were quantified. Average RLU values are normalized as % Activity relative to the respective DMSO control treatment. INDIGO’s Live Cell Multiplex (LCM) Assay was used to quantify cell viability, which is normalized as % Live Cells relative to the DMSO control. Values of ≥ 85% Live Cells at the assay endpoint indicate non-significant cytotoxicity. For both assays, error-bars depict coefficient of variation (%CV). ** and *** = p value ≤ 0.001 and ≤ 0.0001, respectively, as determined by two-way T-test. Reference compounds were obtained from Cayman Chemical, except for TED-347, which was obtained from SelleckChem.

Target Background

The Hippo tumor suppressor pathway coordinates cellular signals that modulate cell proliferation, tissue homeostasis, and organ size. This signaling primarily regulates the ability of YAP, or its paralog TAZ, to bind and co-activate the TEAD family of transcription factors (TEAD1-4).

YAP confers traits that sustain cell proliferation, inhibit apoptosis, promote angiogenesis, and develop resistance to therapies. The TEAD/YAP complex is therefore considered an oncogene regulator, as the dysregulation of YAP is strongly associated with the onset and progression of several cancers, such as prostate and pancreatic cancer. As such, the Hippo pathway is a premier target for the development of novel, specific, small molecule inhibitors.

The evolutionarily conserved Hippo pathway comprises a complex activation cascade of protein kinases. Activated MST1/2 forms a complex with SAV1 to phosphorylate and activate LATS1/2 kinases. Activated LATS in complex with MOB1 phosphorylate YAP and/or TAZ, leading to cytoplasmic retention via association with 14-3-3d. Alternatively, phosphorylated YAP may be ubiquinated and targeted for proteasomal degradation. Under these conditions the expression of genes associated with pro-proliferative cell function is reduced. The inhibition, or dysregulation, of this kinase cascade leads to reduced phosphorylation of YAP, their translocation to the nucleus, and association with TEAD transcription factors. The assembly of an active TEAD/YAP (or TEAD/TAZ) transcription complex induces expression of oncogenic target genes involved in mediating cell proliferation, migration, and survival.

INDIGO’s Hippo pathway assay system utilizes proprietary human cells engineered to provide high-level expression of a hybrid form of the Human Transcriptional Enhanced Associate Domain 4 Protein (TEAD4), whereby the DNA binding domain (DBD) of the native TEAD4 has been substituted with that of the yeast Gal4-DBD. Additionally, these reporter cells express the requisite transcriptional co-activator Human Yes-Associated Protein 1 (YAP) as well as the luciferase reporter gene functionally linked to tandem Gal4 Upstream Activation Sequence (UAS) genetic response elements. The use of a hybrid Gal4(DBD)-TEAD4 is a well-used strategy, and ensures that any treatment-induced changes in reporter gene activity are the direct result of changes in Gal4(DBD)-TEAD4 / YAP transcriptional activity.

To reiterate, the reporter cells provided in INDIGO’s assay present the “Hippo Off,” or oncogenic pathway. Specifically, the Gal4(DBD)-TEAD4 transcription factor and the YAP co-activator are over-expressed to produce a constitutively active transcription complex that delivers high-level expression of the luciferase reporter gene. The primary application of this assay system is to screen test materials for inhibitory activities against the TEAD4 / YAP transcription complex.

Also available as a service

TEAD4/YAP (Hippo Pathway)

Have questions about this assay kit?