FOXO Reporter Kit (PI3K/AKT Pathway)
The FOXO Reporter Kit (PI3K/AKT Pathway) is designed for monitoring the activity of the PI3K (phosphoinositide 3-kinase)/AKT (protein kinase B) signaling pathway and the transcriptional activity of FOXO (forkhead box O) proteins in a cellular model. The kit contains a transfection-ready FOXO3 Expression Vector, a FOXO Luciferase Reporter Vector, which is a PI3K/AKT pathway-responsive reporter, and a Renilla Luciferase Vector. The FOXO Luciferase Reporter Vector contains the firefly luciferase reporter under the control of multimers of the FOXO responsive element located upstream of a minimal promoter. The FOXO reporter vector comes premixed with a constitutively expressing sea pansy (Renilla) luciferase vector, which serves as an 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 also contains the firefly luciferase gene under the control of a minimal promoter, but without the additional FOXO responsive elements. The negative control is critical for determining pathway-specific effects and background luciferase activity.
This product has been cited 14 times.
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Name | Ordering Information |
Mammalian cell line of interest | |
Appropriate cell culture media | |
Lipofectamine™ 2000 Transfection Reagent | Thermo Fisher #11668027 |
Opti-MEM™ I Reduced Serum Medium | Invitrogen #31985-062 |
Clear-bottom, white 96-well tissue culture-treated plate | Corning #3610 |
TWO-Step™ Luciferase (Firefly & Renilla) Assay System | BPS Bioscience #60683 |
Luminometer |
Catalog # | Name | Amount | Storage |
Component A | Reporter Mix (FOXO Luciferase Reporter Vector + constitutively expressing Renilla Luciferase Vector) (60 ng DNA/µl) | 500 µl | -20°C |
Component B | Negative Control Reporter Mix (Non-Inducible Luciferase Vector + constitutively expressing Renilla Luciferase Vector) (60 ng DNA/µl) | 500 µl | -20°C |
Component C | FOXO3 Expression Vector (100 ng DNA/µl) | 250 µl | -20°C |
Component D | Negative Control Expression Vector (Empty Expression Vector, without FOXO3) (100 ng DNA/µl) | 250 µl | -20°C |
Note: These vectors are ready for transient cell transfection. They are NOT SUITABLE for transformation and amplification in bacteria.
The (phosphoinositide 3-kinase)/AKT (protein kinase B) signaling pathway is essential for cell growth and survival. Disruption of this pathway or its regulation has been linked to a variety of cancers and coronary diseases. Mammalian FOXO (forkhead box O) proteins (FOXO1, FOXO3, FOXO4), a subgroup of Forkhead transcription factors, is among the best characterized targets of the PI3K/AKT signaling pathway. These transcription factors function as a trigger for apoptosis by up-regulating genes necessary for cell death. Insulin or growth factors induce activation of PI3K, which in turn activates AKT. AKT directly phosphorylates FOXOs, resulting in the export of FOXOs from the nucleus to the cytoplasm, thereby inhibiting FOXO-dependent transcription.
Essaghir A., et al., 2009 J. Biol. Chem. 284(16):10334-10342
Hennessy B.T., et al., 2005 Nat. Rev. Drug Discov. 4(12):988-1004.