Product Information
Product Usage Information
Cells are lysed in a urea-containing buffer, cellular proteins are digested by proteases, and the resulting peptides are purified by reversed-phase solid-phase extraction. Peptides are then subjected to immunoaffinity purification using a PTMScan® Motif Antibody conjugated to protein A agarose beads. Unbound peptides are removed through washing, and the captured PTM-containing peptides are eluted with dilute acid. Reversed-phase purification is performed on microtips to desalt and separate peptides from antibody prior to concentrating the enriched peptides for LC-MS/MS analysis. CST recommends the use of PTMScan® IAP Buffer #9993 included in the kit.
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Product Description
Background
The MAPK and CDK families of serine/threonine protein kinases play important roles in proliferation and cell cycle control. These kinases phosphorylate threonine or serine residues that are followed by a proline residue (1-3). MAPK phosphorylates substrates with the consensus sequence PX(S/T)P and CDKs phosphorylate substrates containing the consensus sequence (S/T)PXR/K (4,5). Some signaling molecules can be regulated by phosphorylation at a specific threonine followed by an arginine or lysine at the +2 position. For example, conventional PKC isozymes phosphorylate substrates containing a serine or threonine with Arg or Lys at the -3, -2 and +2 positions (6,7). Cell Signaling Technology has developed antibodies that bind to phosphorylation sites containing the T(P/X)R motif for discovery and proteome-wide profiling of kinase substrates.
- Cross, T.G. et al. (2000) Exp Cell Res 256, 34-41.
- Reynolds, C.H. et al. (2000) J Neurochem 74, 1587-95.
- Seger, R. and Krebs, E.G. (1995) FASEB J 9, 726-35.
- Holmes, J.K. and Solomon, M.J. (1996) J Biol Chem 271, 25240-6.
- Songyang, Z. et al. (1996) Mol Cell Biol 16, 6486-93.
- Nishikawa, K. et al. (1997) J Biol Chem 272, 952-60.
- Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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![PTMScan® Phospho-Thr-X-Arg Motif [pT(P/X)R] Kit: Image 1](http://media.cellsignal.cn/product/image/91336_fig02___20210423082221.jpg)
![PTMScan® Phospho-Thr-X-Arg Motif [pT(P/X)R] Kit: Image 2](http://media.cellsignal.cn/product/image/91336_fig03___20210423082231.jpg)