The Wnt/b-catenin signaling pathway controls a large and diverse set of cell fate decisions in embryonic development, adult organ maintenance and disease. Wnt proteins bind to receptors on the cell surface, initiating a signaling cascade that leads to stabilization and nuclear translocation of b-catenin. b-catenin then binds to TCF/LEF transcription factors in the nucleus, leading to transcription and expression of Wnt-responsive genes. Intended Use: Designed for monitoring the activity of Wnt/b-catenin signaling pathway in the cultured cells. Test Principle: The kit contains transfection- ready TCF/LEF luciferase reporter vector, which is a Wnt pathway-responsive reporter. This reporter contains a firefly luciferase gene under the control of multimerized TCF/LEF responsive element located upstream of a minimal promoter. The TCF/LEF reporter is premixed with constitutively expressing Renilla luciferase vector that serves as internal control for transfection efficiency. The kit also includes a non-inducible firefly luciferase vector premixed with constitutively expressing Renilla luciferase vector as negative control. The non- inducible luciferase vector contains a firefly luciferase gene under the control of a minimal promoter, without any additional response elements. The negative control is critical to determining pathway specific effects and background luciferase activity. Kit Components: Reporter (Component A): TCF/LEF luciferase reporter vector + constitutively expressing Renilla luciferase vector. 1x500ul (60ng DNA/ul) Negative Contol Reporter (Component B): Non-inducible luciferase vector + constitutively expressing Renilla luciferase vector. 1x500ul (60ng DNA/ul) Storage and Stability: Store other components at 4C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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