| 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. |
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| Catalog # | 0014-33 |
| 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. |
| CAS Number | N/A |
| Clone Type | Polyclonal |
| Isotype | IgG |
| Host | Rabbit |
| Concentration | 0.35mg/ml |
| Form | Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. |
| Purity | Purified by Protein A and peptide affinity chromatography. |
| Immunogen | Synthetic phospho- (Ser) 14-3-3-binding-motif peptides. |
| 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. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
