FOXO Reporter Kit (PI3K/AKT Pathway)

Catalog #
60643
$565 *
Size: 500 reactions
Qty
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Description

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.

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

Synonyms
FOXO reporter, forkhead reporter, PI3K reporter, AKT reporter, FOXO1 reporter, FOXO3 reporter, FOXO4 reporter, Cell based assay kit, cell-based assay, reporter assay kit
Product Info
Storage and Usage
Citations14
Materials Required But Not Supplied
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  
Format
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.

Background

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.

References

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.