Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-Threonine Antibody (P-Thr-Polyclonal) #9381

Applications Reactivity Sensitivity Source
W IP E-P All Endogenous Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

Specificity / Sensitivity

Phospho-Threonine Antibody (P-Thr-Polyclonal) detects proteins and peptides phosphorylated at threonine residues in a manner largely independent of the surrounding amino acid sequence. The antibody is phospho-specific and may cross-react with some phospho-serine-containing sequences. By ELISA, it recognizes a wide variety of threonine-phosphorylated peptides, and by 2D gel Western blot analysis, it recognizes a large number of presumably threonine-phosphorylated proteins. CST recommends the use of Phospho-Threonine-Proline mAb (p-Thr-Pro-101) #9391 to detect proteins containing threonine followed by proline. (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-Thr-containing peptides. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or calyculin A-treated (0.1 µM for 45 minutes prior to lysis) and subjected to 2-D gel electrophoresis, using Phospho-Threonine Antibody (P-Thr-Polyclonal).

ELISA-Peptide

ELISA-Peptide

Phospho-Threonine Antibody (P-Thr-Polyclonal) ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (T* denotes phosphorylated threonine.)

Background

Much of the dynamic behavior of cellular proteins, including the regulation of molecular interactions (1), subcellular localization (2), and transcriptional regulation (3) is controlled by a variety of post-translational modifications (4). Antibodies specific for these post-translational modifications are invaluable tools in the quest to understand normal and pathogenic molecular and cellular behavior.General protein modification antibodies are designed to react with modified amino acid residues (e.g. phospho-threonine, phospho-tyrosine, acetyl-lysine, nitro-tyrosine) independently of the sequence in which they are embedded. This ability to recognize modified residues in a "context-independent" fashion gives these antibodies broad reactivities, presumably conferring upon them the ability to react with hundreds of distinct proteins. This broad pattern of reactivity makes these antibodies especially valuable in multiplex analyses and target discovery programs.

Protein kinases are among the most abundant eukaryotic regulatory proteins; over 500 separate kinase genes are encoded in mammalian genomes (5,6). In spite of the importance of kinases in eukaryotic biology, relatively few of their physiological targets are known. Phospho-Threonine Antibody (P-Thr-Polyclonal) #9381 and Phospho-Threonine (42H4) mAb #9386 provide powerful tools for discovering targets of serine/threonine kinases, for monitoring and characterizing in vitro threonine phosphorylation reactions as well as for high throughput Ser/Thr kinase drug discovery.

  1. Yaffe, M.B. and Elia, A.E. (2001) Curr Opin Cell Biol 13, 131-8.
  2. Appella, E. and Anderson, C.W. (2001) Eur J Biochem 268, 2764-72.
  3. Jenuwein, T. and Allis, C.D. (2001) Science 293, 1074-80.
  4. Krishna, R.G. and Wold, F. (1993) Adv Enzymol Relat Areas Mol Biol 67, 265-98.
  5. Venter, J.C. et al. (2001) Science 291, 1304-51.
  6. Manning, G. et al. (2002) Science 298, 1912-34.

Application References

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Companion Products

License/Use Restrictions: Use of CST Motif Antibodies within certain methods (e.g., U.S. Patent No.'s 7,198,896 & 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

For Research Use Only. Not For Use In Diagnostic Procedures.

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