CST, 91336S, PTMScan® Phospho-Thr-X-Arg Motif [pT(P/X)R] Kit

PTMScan for studying in the research area. 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. Storage Antibody beads supplied in IAP buffer containing 50% glycerol. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: PTMScan 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. Alternate Names TPR; TXR