Product Pathways - Akt Signaling

Phospho-FoxO1 (Thr24)/FoxO3a (Thr32) Blocking Peptide #1060

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Description

This peptide can used to specifically block Phospho-Fox01 (Thr24)/Fox03a (Thr32) Antibody #9464 reactivity.

Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide has been tested to block Phospho-Fox01 (Thr24)/Fox03a (Thr32) Antibody #9464 signal completely in Western blotting.

Applications

Use as a blocking reagent to evaluate the specifity of antibody reactivity in Western immunoblotting protocols.

Directions for Use

For Western immunoblotting, add 10 µl of antibody and 10 µl of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room temperature for 30 minutes before allowing to react with the blot.

Background

The Forkhead family of transcription factors is involved in tumorigenesis of rhabdomyosarcoma and acute leukemias (1-3). Within the family, three members (FoxO1, FoxO4 and FoxO3a) have sequence similarity to the nematode orthologue DAF-16, which mediates signaling via a pathway involving IGFR1, PI3K and Akt (4-6). Active forkhead members act as tumor suppressors by promoting cell cycle arrest and apoptosis. Increased expression of any FoxO member results in the activation of the cell cycle inhibitor p27Kip1. Forkhead transcription factors also play a part in TGF-β-mediated upregulation of p21CIP1, a process negatively regulated through PI3K (7). Increased proliferation results when forkhead transcription factors are inactivated through phosphorylation by Akt at Thr24, Ser256 and Ser319, which results in nuclear export and inhibition of transcription factor activity (8). Forkhead transcription factors can also be inhibited by the deacetylase sirtuin (SirT1) (9).

1. Anderson, M.J. et al. (1998) Genomics 47, 187-199.
2. Galili, N. et al. (1993) Nat. Genet. 5, 230-235.
3. Borkhardt, A. et al. (1997) Oncogene 14, 195-202.
4. Nakae, J. et al. (1999) J. Biol. Chem. 274, 15982-15985.
5. Rena, G. et al. (1999) J. Biol. Chem. 274, 17179-17183.
6. Guo, S. et al. (1999) J. Biol. Chem. 274, 17184-17192.
7. Seoane, J. et al. (2004) Cell 117, 211-223.
8. Arden, K.C. (2004) Mol. Cell 14, 416-418.
9. Yang, Y. et al. (2005) EMBO J. 24, 1021-1032.

Application References

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Companion Products

• 9464 Phospho-FoxO1 (Thr24)/FoxO3a (Thr32) Antibody