14-3-3 Binding Motif, phosphorylated (Ser)

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.\n\nApplications: \nSuitable for use in, ELISA, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000. By 2D gel Western Blot, recognizes a large number of presumptive 14-3-3 binding proteins.\nImmunoprecipitation: 1:50 \nImmunohistochemistry (Paraffin): 1:200\nELISA: 1:1000. Recognizes a wide range of peptides containing the 14-3-3 binding motif \nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees 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.

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SPECIFICATIONS

Catalog Number

0014-33

Size

100ul

Applications

ELISA, IHC, IP, WB

Hosts

Rabbit

Form

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.

P Type

Pab

Purity

Purified by Protein A and peptide affinity chromatography.

Isotype

IgG

References

(1) Aitken, A. (1995) Trends Biochem. Sci. 20, 95

Additional Info

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.