Forkhead Signaling Antibody Sampler Kit #9946
Product Information
Kit Usage Information
Protocols
- 2599: Western Blotting
- 2880: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow, ChIP Magnetic
- 7074: Western Blotting
- 9461: Western Blotting
- 9464: Western Blotting, Immunoprecipitation (Magnetic)
- 9465: Western Blotting, Immunoprecipitation (Magnetic)
- 9472: Western Blotting
- 12829: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
- 13129: Western Blotting
Product Description
This sampler kit provides an economical means to investigate Forkhead signaling. The kit contains primary and secondary antibodies to perform two Western blots with each antibody.
Specificity / Sensitivity
Phospho-FoxO1 (Thr24)/FoxO3a (Thr32) Antibody detects endogenous levels of FoxO1/FoxO3a only when phosphorylated at Thr24 of FoxO1 or Thr32 of FoxO3a. The antibody cross-reacts with phosphorylated FoxO4 at Thr28, but not with FoxO1 family members phosphorylated at other sites. Phospho-FoxO1 (Ser256) Antibody detects endogenous levels of FoxO1 only when phosphorylated at Ser256. The antibody cross-reacts with Fox04 phosphorylated at Ser193. Phospho-FoxO1 (Thr24)/FoxO3a (Thr32)/Fox04 (Thr28) (4G6) Rabbit mAb detects endogenous levels of FoxO1 when phosphorylated at Thr24, of FoxO3a when phosphorylated at Thr32 or FoxO4 when phosphorylated at Thr28. FoxO1 (C29H4) Rabbit mAb detects endogenous levels of total FoxO1 protein. The antibody does not detect exogenously expressed family members FoxO3a or FoxO4. Phospho-FoxO3a (Ser253) (D18H8) Rabbit mAb detects endogenous levels of FoxO3a only when phosphorylated at Ser253. This antibody may cross-react with FoxO1 when overexpressed and phosphorylated at Ser251 or FoxO4 when overexpressed and phosphorylated at Ser197. Phospho-FoxO3a (Ser318/321) Antibody detects endogenous levels of FoxO3a only when phosphorylated at Ser318/321. The antibody is expected to cross-react with FoxO1 when phosphorylated at Ser322/325 based on the peptide sequence. FoxO3a (D19A7) Rabbit mAb recognizes endogenous levels of total FoxO3a protein. The FoxO4 Antibody detects endogenous levels of FoxO4. The antibody is sensitive to phosphorylation within the antigen and preferrentially detects unphosphorylated FoxO4.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr28 of human Fox04, with a synthetic phosphopeptide corresponding to residues around Ser256 of human Fox01, or with a synthetic peptide corresponding to residues of human Fox04. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with recombinant protein specific to the carboxy terminus of human FoxO3 protein, with a GST-fusion protein corresponding to carboxy-terminal residues of human FoxO1, with a synthetic phosphopeptide corresponding to the sequence of human Fox03a, or with a synthetic phosphopeptide corresponding to residues surrounding Thr28 of human Fox04.
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).
- Anderson, M.J. et al. (1998) Genomics 47, 187-99.
- Galili, N. et al. (1993) Nat Genet 5, 230-5.
- Borkhardt, A. et al. (1997) Oncogene 14, 195-202.
- Nakae, J. et al. (1999) J Biol Chem 274, 15982-5.
- Rena, G. et al. (1999) J Biol Chem 274, 17179-83.
- Guo, S. et al. (1999) J Biol Chem 274, 17184-92.
- Seoane, J. et al. (2004) Cell 117, 211-23.
- Arden, K.C. (2004) Mol Cell 14, 416-8.
- Yang, Y. et al. (2005) EMBO J 24, 1021-32.
限制使用
除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。
专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专
For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit our
Trademark Information page.