Technical Data
0014-33
14-3-3 Binding Motif, phosphorylated (Ser)
Description:
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). This antibody provides a powerful tool for the discovery and characterization of potential 14-3-3 binding proteins containing this motif and for high throughput drug discovery.

Applications:
Suitable for use in, ELISA, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000. By 2D gel Western Blot, recognizes a large number of presumptive 14-3-3 binding proteins.
Immunoprecipitation: 1:50
Immunohistochemistry (Paraffin): 1:200
ELISA: 1:1000. Recognizes a wide range of peptides containing the 14-3-3 binding motif
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ul4°C (-20°C Glycerol)Blue IceRabbit
Concentration:
0.35mg/ml
Immunogen:
Synthetic phospho- (Ser) 14-3-3-binding-motif peptides.
Purity:
Purified by Protein A and peptide affinity chromatography.
Form
Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.
Specificity:
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. Weakly crossreacts with sequences containing phospho-Thr instead of phospho-Ser in this motif; with sequences containing phospho-Ser surrounded by Phe at the +1 position; with Arg/Lys at the -3 position. No crossreactivity is observed with corresponding non-phosphorylated sequences or with other phospho- Thr/Ser/Tyr -containing motifs.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
(1) Aitken, A. (1995) Trends Biochem. Sci. 20, 95–97. (2) Zha, J. et al. (1996) Cell 87, 619–628. (3) Piwnica-Worms, H. (1999) Nature 401, 535–537. (4) Tzivion, G. et al. (1998) Nature 394, 88–92. (5) Xing, H. (2000) EMBO J. 19, 349–358. (6) Muslin, A.J. (1996) Cell 84, 889–897. (7) Yaffe, M.B. et al. (1997) Cell 91, 961–971.