Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb #9606

Applications Reactivity Source Isotype
W IP E-P All Mouse IgG1

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

Specificity / Sensitivity

Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb binds peptides and proteins containing phospho-Ser surrounded by Pro at the +2 position and Arg/Lys at the -3 position. By ELISA, the antibody recognizes a wide range of peptides containing this phosphorylated 14-3-3 binding motif in a manner that is phospho-specific and largely independent of the surrounding amino acid sequence. 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-Ser/Thr/Tyr containing motifs. Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb complements our polyclonal Phospho-(Ser) 14-3-3 Binding Motif Antibody #9601 by showing slightly different and overlapping specificity. (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

Monoclonal antibody is produced by immunizing mice with phospho-(Ser) 14-3-3 binding motif peptides (KLH-coupled).

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or calyculin A-treated, using Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb (left) or Phospho-(Ser) 14-3-3 Binding Motif Antibody #9601 (right).

ELISA-Peptide

ELISA-Peptide

Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb DELFIA® assay: Signal-to-noise ratio of phospho- versus nonphospho-14-3-3 binding motif peptides. (S* denotes phosphorylated serine.) (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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

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Companion Products

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.

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