Product Pathways - Motif Antibodies

Phospho-(Ser) 14-3-3 Binding Motif Antibody #9601

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Applications	Reactivity	Source
W IP IHC-P E-P	All	Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected

Specificity / Sensitivity

Phospho-(Ser) 14-3-3 Binding Motif Antibody binds peptides and proteins containing a motif composed of phospho-Ser with proline at the +2 position and arginine or lysine at the -3 position. Antibody binding is phospho-specific and largely independent of other surrounding amino acids. The antibody weakly cross-reacts with sequences containing phospho-Thr instead of phospho-Ser in this motif, and with sequences containing phospho-Ser surrounded by Phe at the +1 position and Arg/Lys at the -3 position. No cross-reactivity is observed with corresponding nonphosphorylated sequences or with other phospho-Thr/Ser/Tyr-containing motifs. By ELISA this antibody recognizes a wide range of peptides containing the 14-3-3 binding motif, and by 2D gel Western blot analysis it recognizes a large number of presumptive 14-3-3 binding proteins. (U.S. Patent No's.: 6,441,140; 6,982,318; 7,259,022; 7,344,714; U.S.S.N. 11,484,485; and all foreign equivalents.)

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic phospho-(Ser) 14-3-3-binding-motif containing peptides (KLH-coupled). Antibodies are purified by protein A and peptide affinity chromatography.

Background

The 14-3-3 proteins are a highly conserved family of proteins involved in the regulation of cell survival, apoptosis, proliferation and checkpoint control (1-5). Biological regulation by 14-3-3 is mediated through phosphorylation-dependent protein-protein interactions (6). Two different phospho-Ser-containing motifs are found within nearly all known 14-3-3 binding proteins (7). Motif 1 (Arg/Lys and Ser at positions -3 and -2, phospho-Ser at position 0, and Pro at position +2) is found in critical regulatory proteins including Bad, cdc25C, FKHRL1, PKC and c-Raf (5,7). Phospho-(Ser) 14-3-3 Binding Motif Polyclonal and (4E2) Monoclonal Antibodies provide powerful tools for the discovery and characterization of potential 14-3-3 binding proteins containing this motif and for high throughput drug discovery.

1. Aitken, A. (1995) Trends Biochem Sci 20, 95-7.
2. Zha, J. et al. (1996) Cell 87, 619-28.
3. Piwnica-Worms, H. (1999) Nature 401, 535, 537.
4. Tzivion, G. et al. (1998) Nature 394, 88-92.
5. Xing, H. et al. (2000) EMBO J 19, 349-58.
6. Muslin, A.J. et al. (1996) Cell 84, 889-97.
7. Yaffe, M.B. et al. (1997) Cell 91, 961-71.

Application References

• Diao, J. et al. (2001) X protein of hepatitis B virus inhibits Fas-mediated apoptosis and is associated with up-regulation of the SAPK/JNK pathway. J. Biol. Chem. 276 (11), 8328-8340. This article references the use of Phospho-(Ser) 14-3-3 Binding Motif Antibody in the following applications: Western Blot
• Shen, Y. H. et al. (2003) Significance of 14-3-3 self-dimerization for phosphorylation-dependent target binding. Mol. Biol. Cell 14, 4721-4733. This article references the use of Phospho-(Ser) 14-3-3 Binding Motif Antibody in the following applications: Western Blotting
• Liu, M. Y. et al. (2002) 14-3-3 interacts with the tumor suppressor tuberin at Akt phosphorylation site(s). Cancer Res. 62, 6475-6480. This article references the use of Phospho-(Ser) 14-3-3 Binding Motif Antibody in the following applications: Western Blotting
• Brunet, A. et al. (2002) 14-3-3 transits to the nucleus and participates in dynamic nucleocytoplasmic transport. J. Cell Biol. 156 (5), 817-828. This article references the use of Phospho-(Ser) 14-3-3 Binding Motif Antibody in the following applications: Western Blotting
• Zang, M. et al. (2001) Microtubule integrity regulates Pak leading to Ras-independent activation of Raf-1. J. Biol. Chem. 276 (27), 25157-25165. This article references the use of Phospho-(Ser) 14-3-3 Binding Motif Antibody in the following applications: Western Blot
• Foschi M, F. et al. (2001) Endothelin-1 induces serine phosphorylation of the adaptor protein p66Shc and its association with 14-3-3 protein in glomerular mesangial cells. J. Biol. Chem. 276 (28), 26640-26647. This article references the use of Phospho-(Ser) 14-3-3 Binding Motif Antibody in the following applications: Western Blot

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

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License/Use Restrictions: Use of CST Motif Antibodies within certain methods (e.g., U.S. Patent No.'s 7,198,896 & 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commerical licensing terms please contact CST Business Development at cbunker@cellsignal.com.