Transfection Collection™ : TCF/LEF Transient Pack Wnt / β-catenin Signaling Pathway

Catalog #
79273
$750 *
Size: 100 reactions
Qty
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Description

The TCF/LEF Transient Pack is designed to provide the tools necessary for transiently transfecting and monitoring the activity of Wnt / b-catenin signaling pathway in the cultured HEK293 cells. The TCF/LEF Transient Pack contains transfection-ready vectors containing firefly luciferase as a Wnt pathway-responsive reporter and constitutively expressing Renilla luciferase as a transfection control.  It also includes the TWO-Step Luciferase detection reagents to detect both luciferase activities and specialized medium for growing and assaying HEK293 cells.

The key to the TCF/LEF Transient Pack is the 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 pack 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.

Additionally, the pack includes cell culture medium (BPS Medium 1) that has been optimized for use with HEK293 cells*. BPS Medium 1 includes MEM medium, 10% fetal bovine serum, 1% non-essential amino acids, sodium pyruvate, and 1% Pen/Strep.   Finally, the pack provides the TWO-Step Luciferase (Firefly & Renilla) Assay System. These luciferase reagents provide highly sensitive, stable detection of firefly luciferase activity and Renilla luciferase activity.  The TWO-step luciferase reagents can be used directly in cells in growth medium, and can be detected with any luminometer; automated injectors are not required.

*Note:  the kit may be used with other cell lines than HEK293, but an alternate cell culture medium may be required for optimal cell growth.

This product has been cited 8 times.

Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.

Synonyms
WNT reporter, TCF reporter, LEF reporter, luciferase reporter
Product Info
Storage and Usage
Citations8
Format

Concentration
Both the Reporter and Negative Control Reporter are provided at 60 ng DNA/µl (500 µl each)
UniProt #
P36402
Background

The Wnt / β-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 β-catenin. β-catenin then binds to TCF/LEF transcription factors in the nucleus, leading to transcription and expression of Wnt-responsive genes.

References
Chen B et al. (2009) Small molecule-mediated disruption of Wnt-dependent signaling in tissue regeneration and cancer. Nature Chemical Biology 5(2): 100-107.