Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-Threonine-X-Arginine Antibody #2351

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

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

Specificity / Sensitivity

Phospho-Threonine-X-Arginine Antibody detects endogenous levels of proteins containing the phospho-Thr-X-Arg motif. This antibody detects phosphorylated Thr followed by Arg or Lys at the +2 position, though its reactivity is lower for Lys compared to Arg at the +2 position. The antibody does not cross-react with nonphospho-Thr or phospho-Ser in the same motif. It recognizes phospho-Thr in the FFT*R motif in PKCbeta II but does not recognize phospho-Thr in other motifs that lack Lys or Arg at +2. (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 a synthetic peptide (KLH-coupled) containing the phospho-Thr-X-Arg motif. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated, TPA-treated or calyculin A-treated, using Phospho-Threonine-X-Arginine Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing staining of proteins containing phosphorylated threonine-X-arginine motifs, using Phospho-Threonine-X-Arginine Antibody.

ELISA-Peptide

ELISA-Peptide

Phospho-Threonine-X-Arginine Antibody ELISA: Signal-to-noise ratio of phospho- versus nonphospho-peptides containing the phospho-threonine-X-arginine motif. (T* denotes phosphorylated threonine.)


Background

Some signaling molecules can be regulated by phosphorylation at a specific threonine followed by arginine or lysine at the +2 position. For example, conventional PKC isozymes phosphorylate substrates containing serine or threonine with Arg or Lys at the -3, -2 and +2 positions (1-2). c-Raf, a mitogen-activated protein kinase and the main effector recruited by GTP-bound Ras, is phosphorylated at Thr481 and Thr491 followed by Lys at the +2 position (3). Phosphorylation of these sites is important for enzyme activities. To determine the phosphorylation state of Thr in the Thr-X-Arg motif, and to identify potential new phosphorylation sites with this motif, CST has developed a Phospho-Threonine X-Arginine Antibody that recognizes phosphorylated Thr followed by Arg or Lys at the +2 position.

  1. Nishikawa, K. et al. (1997) J Biol Chem 272, 952-60.
  2. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
  3. Zhang, B.H. and Guan, K.L. (2000) EMBO J 19, 5429-39.

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

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