Phospho-(Ser) 14-3-3 Binding Motif Antibody 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. 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-Thr/Ser/Tyr-containing motifs. By ELISA this antibody recognizes a wide range of peptides containing the 14-3-3 binding motif, and by 2D gel Western blot analysis it recognizes a large number of presumptive 14-3-3 binding proteins. (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
Polyclonal antibodies are produced by immunizing animals with synthetic phospho-(Ser) 14-3-3-binding-motif containing peptides . Antibodies are purified by protein A and peptide affinity chromatography.
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.
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