FOXO Reporter Kit (PI3K/AKT Pathway)

Catalog #: 60643
Size: 500 reactions
$525
Background:
The PI3K/AKT 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 protein (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 upregulating 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.
Description:
The FOXO Reporter kit is designed to monitor activity of the PI3K/AKT signaling pathway and the transcriptional activity of FOXO proteins in cultured cells. The kit contains the transfection-ready FOXO3 expression vector and the FOXO luciferase reporter vector, which is a PI3K/Akt pathway-responsive reporter. This reporter contains a firefly luciferase gene under the control of multimers of the FOXO responsive element located upstream of a minimal promoter. The FOXO reporter is premixed with a constitutively-expressing Renilla (sea pansy) luciferase vector, which serves as an internal control for the 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 any additional response elements. The negative control is critical to determining pathway-specific effects and background luciferase activity.
Synonym(s):
FOXO reporter, forkhead reporter, PI3K reporter, AKT reporter, FOXO1 reporter, FOXO3 reporter, FOXO4 reporter
Supplied As:
Reporter: FOXO luciferase reporter vector + constitutively-expressing Renilla luciferase vector
Negative Control Reporter: Non-inducible luciferase vector + constitutively-expressing Renilla luciferase vector
Biological Activity:

This cell line is validated for response to stimulation by Forskolin and to the treatment with an inhibitor of the cAMP/PKA signaling pathway.

Concentration:

The Reporter and Negative Control Reporter are provided at 60 ng DNA/µl (500 µl each). The FOXO3 Expression Vector and Negative Control Expression Vector are provided at 100 ng DNA/µl (250 µl each).

Instructions for use:
See data sheet for detailed transfection and assay protocols.
Storage / Stability:

Stable at least 12 months from date of receipt, when stored as directed (-20°C)

Application(s):
• Monitor PI3K/AKT signaling pathway activity and FOXO transcriptional activity.
Reference(s):

1. Essaghir, A., et al. (2009). J. Biol. Chem. 284(16):10334-10342
2. Hennessy B.T., et al. (2005). Nat. Rev. Drug Discov. 4(12):988-1004.

Application Reference(s):
1. Barley sprout extracts reduce hepatic lipid accumulation in ethanol-fed mice by activating hepatic AMP-activated protein kinase (2017)
2. FOXO responses to Porphyromonas gingivalis in epithelial cells (2015)

Notes:
The vectors are suitable for transient transfection. They are NOT suitable for transformation and amplification in bacteria.
Warning(s):
Avoid freeze/thaw cycles
Scientific Category:
Cell Signaling Pathway
Product Type:
Reporter Assay Kit
Data shown is lot-specific. Contact us for specific information on other lots.

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Product Datasheets

60643.pdf