Product Pathways - Motif Antibodies

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

• pathway
• more info
• application references
• datasheet PDF
• MSDS PDF
• protocols

Applications	Reactivity	Source
W IP IHC-P E-P	All	Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected

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 rabbits with synthetic phospho-Thr-containing peptides (KLH-coupled). Antibodies are purified by protein A and peptide affinity chromatography.

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 posttranslational modifications (4). Antibodies specific for these posttranslational 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. (2000) Science 291, 1304-1351.
6. Manning, G. et al. (2002) Science 298, 1912-1934.

Application References

• Haldrup, A. et al. (2000) Down-regulation of the PSI-F subunit of Photosystem I (PSI) in Arabidopsis thaliana. Journal of Biological Chemistry 275, 31211-31218. This article references the use of Phospho-Threonine Antibody (P-Thr-Polyclonal) in the following applications: Western Blot
• Ma, T. et al. (2000) Cell cycle-regulated phosphorylation of p220(NPAT)by cyclin E/Cdk2 in Cajal bodies promotes histone gene transcription. Genes and Development 14, 2298-2313. This article references the use of Phospho-Threonine Antibody (P-Thr-Polyclonal) in the following applications: Western Blotting
• Scheinfeld, M. H. et al. (2003) Amyloid beta protein precursor is phosphorylated by JNK-1 independent of, yet facilitated by, JNK-interacting protein (JIP)-1. J. Biol. Chem. 278, 42058-42063. This article references the use of Phospho-Threonine Antibody (P-Thr-Polyclonal) in the following applications: Western Blotting
• Wada, S. et al. (2005) Phosphorylation of candida glabrata ATP-binding cassette transporter Cdr1p regulates drug efflux activity and ATPase stability. J. Biol. Chem. 280, 94-103. This article references the use of Phospho-Threonine Antibody (P-Thr-Polyclonal) in the following applications: Western Blotting
• Gronborg, M. et al. (2002) A mass spectrometry-based proteomic approach for identification of serine/threonine-phosphorylated proteins by enrichment with phospho-specific antibodies: identification of a novel protein, Frigg, as a protein kinase A substrate. Mol. Cell. Proteomics 1, 517-527. This article references the use of Phospho-Threonine Antibody (P-Thr-Polyclonal) in the following applications: IP Western Blotting
• Aoyama, K. et al. (2001) Molecular cloning and characterization of a novel dual specificity phosphatase, LMW-DSP2, that lacks the cdc25 homology domain. J. Biol. Chem. 276, 27575-27583. This article references the use of Phospho-Threonine Antibody (P-Thr-Polyclonal) in the following applications: Western Blotting

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

• 9386 Phospho-Threonine (42H4) Mouse mAb
• 9391 Phospho-Threonine-Proline Mouse mAb (P-Thr-Pro-101)
• 9411 Phospho-Tyrosine Mouse mAb (P-Tyr-100)
• 7900 PhosphoScan® Kit (P-Tyr-100)
• 9416 Phospho-Tyrosine Mouse mAb (P-Tyr-102)
• 9614 Phospho-Akt Substrate (RXRXXS/T) (110B7E) Rabbit mAb
• 9624 Phospho-PKA Substrate (RRXS/T) (100G7E) Rabbit mAb
• 9634 Phospho-(Ser/Thr) PDK1 Docking Motif (18A2) Mouse mAb
• 7071 Phototope^®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

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 commerical licensing terms please contact CST Business Development at cbunker@cellsignal.com.