Myc Reporter Kit (Myc Signaling Pathway)
The Myc Pathway Reporter kit is designed for monitoring the activity of the Myc signaling pathway in cultured cells. The kit contains a transfection-ready expression vector 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 kit 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. The negative control is critical for determining pathway-specific effects and background luciferase activity.
For assays in more difficult to transfect cell types, we suggest using our lentiviral products. The corresponding lentiviral product for this kit is BPS Bioscience #78628.
This product has been cited 3 times.
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- Mammalian cell line and appropriate cell culture medium
- 96-well tissue culture plate or 96-well tissue culture treated white clear-bottom assay plate (Corning # 3610)
- Transfection reagents for mammalian cell line. We use Lipofectamine™ LTX with PLUSTM reagent (Life Technologies #15338100) or Lipofectamine™ 2000 (Life Technologies # 11668027). However, other transfection reagents should work equally well.
- Opti-MEM I Reduced Serum Medium (Life Technologies #31985-062)
- TWO-Step Luciferase (Firefly & Renilla) Assay System (BPS Bioscience #60683): This system assays cells directly in growth medium. It can be used with any luminometer. Automated injectors are not required.
- Luminometer
Component | Specification | Amount | Storage |
Reporter (Component A) | Myc luciferase reporter vector + constitutively-expressing Renilla luciferase vector | 500 µl (60 ng DNA/µl) |
-20°C |
Negative Control Reporter (Component B) | Non-inducible luciferase vector + constitutively-expressing Renilla luciferase vector | 500 µl (60 ng DNA/µl) |
-20°C |
c-Myc Expression Vector (Component C) | c-Myc expression vector | 2 x 250 µl (100 ng DNA/µl) |
-20°C |
Negative Control Expression Vector (Component D) | Empty expression vector without c-Myc | 2 x 250 µl (100 ng DNA/µl) |
-20°C |
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.
Pelengaris S, et al. (2002) c-MYC: more than just a matter of life and death. Nat. Rev. Cancer. 2(10): 764-76.