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LEADING LIGHT® Wnt Cell Line Assay Reagents

For use with the LEADING LIGHT® Wnt Reporter Cell Line
 
ENZ-60004-0001 1 Pack 420.00 USD
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  • Can be used as a supplement to the Wnt LEADING LIGHT® Wnt Reporter Assay Starter Kit (Prod. No. ENZ-61001)
  • When used in the Wnt LEADING LIGHT® Wnt Reporter Assay Starter Kit, there are many advantages of using this line of products:
    • True end-point detection system
    • High sensitivity (Wnt3a EC50 = 45.9 ng/ml)
    • Excellent reproducibility (Z'-factor of 0.74)
    • High signal-to-noise ratio without the need for Lithium Chloride to boost the signal
    • High-throughput screening applications (including 384-well microplates)
The LEADING LIGHT® Wnt Cell Line Assay Reagents are intended for use with the LEADING LIGHT® Wnt Reporter Cell Line, which is the basis for a cell-based luciferase activity test ENZ-61001 LEADING LIGHT® Wnt Reporter Assay Starter Kit suitable for a 96-well plate format. The system contains an engineered 3T3 mouse fibroblast cell line, which expresses the firefly luciferase reporter gene under the control of Wnt-responsive promoters (TCF/LEF). The luciferase activity from the reporter gene in this cell line can be up-regulated in a dose-dependent manner upon the addition of exogenous Wnt protein/Wnt agonist or down-regulated by a further addition of a Wnt antagonist to the cell culture medium. This system can be used to elucidate the functions/activities of different Wnt-related ligands such as Wnt, DKK, etc. This system can also be used for screening small molecules and antibodies for their ability to act as Wnt inhibitors or Wnt agonists.
 
The assay has been used successfully in different assay formats (including HTS applications) to identify several distinct categories of small molecule compounds that modulate the Wnt signaling pathway.

The LEADING LIGHT® Wnt Cell Line Assay Reagents contains Luciferase substrate and assay buffer, Lithium Chloride, HEPES, Wnt3a protein (positive control), Dkk-1 protein, and a 96-well black plate.

Note: the complete LEADING LIGHT® Wnt Reporter Assay Starter Kit (Prod. No. ENZ-61001) is available.

Product Specification

Alternative Name:Wnt reporter reagents
 
Shipping:Shipped on Dry Ice
 
Short Term Storage:-80°C
 
Long Term Storage:-80°C
 
Use/Stability:Upon receipt, store at –80°C. When stored properly, these reagents are stable for one year from date received.
 
Scientific Background:Wnt ligands bind to Frizzled (Fz) and LRP5/6 receptors to trigger a signaling cascade that leads to stabilization of beta-catenin, which can enter into the nucleus to form a complex with T cell transcription factor (TCF/LEF) to activate Wnt target gene expression. Canonical Wnt signaling is required for embryo-genesis and adult tissue maintenance and is involved in tumorigenesis and development of many human degenerative diseases. Studies relating to Wnt signaling have advanced research in molecular embryology, stem cell biology, tumorigenesis, regenerative medicine, and rational drug discovery.
 
ENZ-61001 wnt inhib
Reporter cells were treated with the indicated doses of Dkk-1 in the presence of 200 ng/mL (in-well) Wnt3a. After Dkk-1 treatment, luciferase activity in the cells was measured. Dkk-1 inhibits Wnt3a-elevated luciferase levels in a dose dependent manner.
ENZ-61001 Fig1
Schematic of the canonical Wnt signaling pathway and its components. Luciferase activity from the reporter gene in the LEADING LIGHT® Wnt reporter cell line can be up- or down-regulated in a dose-dependent manner upon the addition of exogenous Wnt protein/Wnt agonist or Wnt antagonist (Dkk) to the cell culture medium.
ENZ-61001 wnt activ Li Cl
Activation of Wnt reporter gene by LiCl in the Wnt Reporter cell system. Cells were treated with the indicated doses of LiCl. After LiCl treatment, luciferase activity was measured. The chemiluminescence in the LiCl-treated cells increased in a dose-dependent fashion.
ENZ-61001 wnt activ
Wnt reporter cells were treated with indicated doses of Wnt3a protein. After Wnt3a treatment, the luciferase activity assay was performed. The chemiluminescence in the Wnt3a-treated cells increased in a dose-dependent manner.
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ENZ-61001 wnt inhib ENZ-61001 Fig1 ENZ-61001 wnt activ Li Cl ENZ-61001 wnt activ

Product Literature References

ARID3B directly regulates ovarian cancer promoting genes: A. Bobbs, et al.; PLoS One 10, e0131961 (2015), Abstract; Full Text
Cardiomyocyte differentiation of pluripotent stem cells with SB203580 analogues correlates with Wnt pathway CK1 inhibition independent of p38 MAPK signaling: F. Laco, et al.; J. Mol. Cell. Cardiol. 80, 56 (2015), Abstract;
Chemical and genetic evidence for the involvement of Wnt antagonist Dickkopf2 in regulation of glucose metabolism: X. Li, et al.; PNAS 109, 11402 (2012), Abstract; Full Text

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