Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-Threonine (42H4) Mouse mAb #9386

Applications Reactivity Sensitivity Isotype
W IP E-P All Endogenous Mouse IgM

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 (42H4) Mouse mAb binds phosphorylated threonine residues in a manner largely independent of the surrounding amino acid sequence. The antibody is phospho-specific but does not cross-react with phospho-tyrosine-containing sequences. It does show slight cross-reactivity with a few phospho-serine-containing peptides. By ELISA, it recognizes a wide variety of threonine-phosphorylated peptides. (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

Monoclonal antibody is produced by immunizing animals with phospho-Thr-containing peptides.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or treated with calyculin A, a threonine phosphatase inhibitor, or extracts from NIH/3T3 cells, untreated or treated with pervanadate, a tyrosine phosphatase inhibitor, using Phospho-Threonine (42H4) Mouse mAb (upper) or Phospho-Tyrosine Monoclonal Antibody (P-Tyr-100) #9411 (lower).

ELISA-Peptide

ELISA-Peptide

Phospho-Threonine (42H4) Mouse mAb 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) Monoclonal Antibody #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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