Transfection Collection™ : Myc Transient Pack (Myc Signaling Pathway)
The Myc Transient Pack is designed to provide the tools necessary for transiently transfecting and monitoring the activity of the Myc signaling pathway in cultured HCT116 cells. The Myc Transient Pack contains transfection-ready vectors containing firefly luciferase as a Myc 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 HCT116 cells.
The key to the Myc Transient Pack is the expression vectors for c-Myc and Myc luciferase reporter vector. Inside the cells, c-Myc will bind to Max, translocate to the nucleus, and induce expression of the Myc luciferase reporter vector. This reporter contains the firefly luciferase gene under the control of multimerized Myc responsive elements located upstream of a minimal promoter. The Myc reporter is premixed with constitutively-expressing Renilla (sea pansy) luciferase vector, which serves as an internal positive control for transfection efficiency.
The pack also includes a non-inducible firefly luciferase vector premixed with constitutively-expressing Renilla luciferase vector as a negative control. The non-inducible luciferase vector contains a firefly luciferase gene under the control of a minimal promoter, but without any additional response elements. This negative control is critical for determining pathway-specific effects and background luciferase activity. An “empty” expression vector without the c-Myc gene is also provided as an additional negative control.
Additionally, the pack includes cell culture medium that has been optimized for use with HCT116 cells*. HCT116 is a human colon cancer cell line with a mutated β -catenin which leads to the accumulation of β-catenin and constitutive activation of the Myc signaling pathway. This medium includes 10% fetal bovine serum, and 1% Pen/Strep. Finally, the pack provides the TWO-Step Luciferase (Firefly & Renilla) Assay System. These 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 HCT116, but an alternate cell culture medium may be required for optimal cell growth. For HEK293 cells, we recommend using BPS Medium 1 (BPS Bioscience, #79259).
Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.
The Myc signaling pathway plays an important role in cell proliferation, differentiation, transformation and apoptosis. The c-Myc protein is a transcription factor that heterodimerizes with Max to regulate Myc signaling pathway responsive genes. Myc mutations have been linked to the development of a number of human cancers, including Burkitt’s lymphoma, cervical, ovarian, breast, lung and pancreatic carcinoma, making Myc a promising therapeutic target for cancer treatment.
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