Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

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

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R Mk Endogenous 78 to 82, 95 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-Fox01 (Thr24)/Fox03a (Thr32) Antibody detects endogenous levels of Fox01/Fox03a only when phosphorylated at threonine 24 of Fox01 or threonine 32 of Fox03a. The antibody cross-reacts with phosphorylated Fox04 at threonine 28, but not with Fox01 family members phosphorylated at other sites.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr28 of human Fox04. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HT29 cells, serum starved or serum treated, using Phospho-Fox01 (Thr24)/Fox03a (Thr32) Antibody.

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 p27 Kip1. Forkhead transcription factors also play a part in TGF-β-mediated upregulation of p21 Cip1, 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.
  10. Camper-Kirby, D. et al. (2001) Circ Res 88, 1020-7.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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