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TCF/LEF Reporter

Luciferase  GFP Lentiviral luciferase Lentiviral GFP
 
Cignal TCF/LEF Reporter (GFP) Kit: CCS-018G
The TCF/LEF Reporter kit is designed to monitor the activity of Wnt signal transduction pathways in cultured cells. The kit contains transfection-ready TCF/LEF reporter construct as well as positive and negative controls. The Wnt signaling pathway leads to the dephosphorylation, stabilization, and nuclear translocation of β-catenin. The stabilized β-catenin complexes with the TCF/LEF transcription factors, leading to the activation of Wnt-responsive genes. The TCF/LEF reporter encodes the Monster GFP gene under the control of a minimal (m)CMV promoter and tandem repeats of the TCF/LEF transcriptional response element (TRE). We have experimentally optimized the number of response elements as well as the intervening sequence between response elements to maximize the signal to noise ratio.

Using a simple GFP reporter assay, the TCF/LEF Reporter (GFP) can easily and rapidly:

  • Monitor Wnt signaling pathway activity in cells
  • Study chemical compounds or drugs
  • Analyze phenotypes of RNAi or gene over expression

See the complete transcription factor reporter assay list.

 

Kit Components How It Works Manual & Resources Related products
 
10-Pathway Arrays
Component Specification Concentration (total volume)
TCF/LEF Reporter An inducible TCF/LEF-responsive GFP reporter (100 ng/µl; 500 µl)
Negative control  A non-inducible GFP reporter (100 ng/µl; 500 µl)
Positive control  A constitutively expressing GFP reporter (100 ng/µl; 250 µl)
 

Kit Components How It Works Manual & Resources Related Products
 
  • Cignal Technology Overview

  • Brief Protocol

The Cignal Reporter Assays (GFP) include pre-formulated, transfection-ready reporter, negative control, and positive control. The transcription factor reporter and negative control are transfected and subjected to experimental treatments, in parallel. GFP expression is quantitated using a flow cytometer, fluorescent microscope, or fluorometer. The change in the activity of each signaling pathway is determined by comparing the GFP activities in treated versus untreated transfectants. The identically treated negative control transfectants serve as a specificity control. The positive control serves as a control for transfection efficiency, by monitoring GFP expression from the constitutively expressing CMV-GFP reporter.

  • Performance Data

Excellent signal to noise ratio

Fluorescence Microscopy: Cignal TCF/LEF-GFP reporter measures activation of the Wnt signaling pathway

293-H cells were transfected with the Cignal TCF/LEF-GFP reporter or negative control (for transfection protocol, refer to our user manual). After 16 hours of transfection, medium was changed to assay medium (Opti-MEM + 0.5% FBS + 0.1mM NEAA + 1mM Sodium pyruvate + 100 U/ml penicillin + 100 µg/ml streptomycin). After 24 hours of transfection, cells were treated with 50mM LiCl. After 18 hours of treatment, bright field and fluorescent images were taken of the cultures transfected with the negative control reporter (A and B, respectively) and the Cignal TCF/LEF-GFP reporter (C and D, respectively).

Quantitative Fluorometry: Cignal TCF/LEF-GFP reporter measures activation of the Wnt signaling pathway

293-H cells were transfected with the Cignal TCF/LEF-GFP reporter or negative control (for transfection protocol, refer to our user manual). After 16 hours of transfection, medium was changed to assay medium (Opti-MEM + 0.5% FBS + 0.1mM NEAA + 1mM Sodium pyruvate + 100 U/ml penicillin + 100 µg/ml streptomycin). After 24 hours of transfection, cells were treated with 50mM LiCl. After 18 hours of treatment, medium was removed from the wells and fluorescence activity was measured using a fluorometer. The fluorescent activity present in treated and non-treated negative control wells was subtracted from the fluorescent activity in treated and non-treated Cignal TCF/LEF-GFP reporter wells and relative fluorescence activities are expressed as arbitrary units. Experiments were done in triplicates, and the standard deviations are indicated.

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Kit Components How It Works Manual & Resources Related Products
 
User Manuals Cignal Reporter Assay Kit User Manual (PDF)
White Paper Cignal Reporter Assay Kit: A High Performance Tool for Assessing the Functions of Genes, Biologics and Small Molecule Compounds (PDF)
Web Seminar Attend a live on-line seminar hosted by an Application Scientist about Cignal Reporters
Technical Brochure Cignal Reporter Assay Kits Technical Brochure (PDF)
Power Point Cignal™ Cell-Based Assays for Rapidly Analyzing Signaling Pathway Activity (PDF)
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Kit Components How It Works Manual & Resources Related Products
 
Pathway reporters Complete list of signaling pathway reporters
Attractene Transfection Achieve high transfection efficiencies and minimal cytotoxicity in your transfections
10-Pathway Arrays Simultaneously measure the activities of ten (10) signaling pathways in a single experiment
shRNA Plasmids Pathway related gene knockdowns - Guaranteed 70% gene knockdown

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