Phospho-(Ser) 14-3-3 Binding Motif Antibody #9601
Filter:
- WB
- IP
- IHC
Western blot analysis of extracts from 293 cells transfected with Bad or Bad Ser112 and/or Ser136 mutant fusion proteins, using Phospho-(Ser) 14-3-3 Binding Motif Antibody (top), Phospho-Bad (Ser136) Antibody #9295 (middle) or Phospho-Bad (Ser112) Antibody #9291 (bottom).
Supporting Data
| REACTIVITY | All |
| SENSITIVITY | Endogenous |
| MW (kDa) | |
| SOURCE | Rabbit |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- IHC-Immunohistochemistry
Species Cross-Reactivity Key:
- All-All Species Expected
- Related Products
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Immunohistochemistry (Paraffin) | 1:100 |
| Peptide ELISA (DELFIA) | 1:500 |
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
Protocol
Specificity / Sensitivity
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.
Species Reactivity:
All Species Expected
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.
- Aitken, A. (1995) Trends Biochem Sci 20, 95-7.
- Zha, J. et al. (1996) Cell 87, 619-28.
- Piwnica-Worms, H. (1999) Nature 401, 535, 537.
- Tzivion, G. et al. (1998) Nature 394, 88-92.
- Xing, H. et al. (2000) EMBO J 19, 349-58.
- Muslin, A.J. et al. (1996) Cell 84, 889-97.
- Yaffe, M.B. et al. (1997) Cell 91, 961-71.
限制使用
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For Research Use Only. Not For Use In Diagnostic Procedures.
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