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).
Suitable for use in ELISA, Western Blot and Immunoprecipitation. Other applications not tested.
Peptide ELISA: 1:1000
Western Blot: 1:4000
Immunoprecipitation: 1:20 (Denatured proteins).
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
| Not determined|
|Synthetic phospho-(Ser) 14-3-3 binding motif peptides (KLH).|
|Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 0.02% sodium azide, 50% glycerol.|
|Recognizes peptides/proteins containing phospho-Ser surrounded by Pro at the +2 position and Arg/Lys at the -3 position. By ELISA, 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. Weakly crossreacts 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 crossreactivity is observed with corresponding nonphosphorylated sequences.|
|Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.|
1. Aitken, A., Trends Biochem. Sci. 20: 95-97 (1995). 2. Zha, J., et al., Cell 87: 619–628 (1996). 3. Piwnica-Worms, H., Nature 401: 535-537 (1999). 4. Tzivion, G., et al., Nature 394: 88-92 (1998). 5. Xing, H., EMBO J. 19: 349-358 (2000). 7. Muslin, A.J., Cell 84: 889-897 (1996). 7. Yaffe, M.B., et al., Cell 91: 961-971 (1997).|