The motif logo was generated from a PhosphoScan® LC-MS/MS experiment using 471 nonredundant tryptic peptides derived from Jurkat cells treated with Calyculin A #9902 and pervanadate, and immunoprecipitated using Immunoaffinity beads from PTMScan® Phospho-CK2 Substrate Motif (S*/T*DXE) Kit #12170. This PhosphoSitePlus® (PSP) logo represents the enrichment of an amino acid at a residue position relative to the abundance of that amino acid in the background, in this case, the human proteome. Residues represented at the top of the logo are the most enriched, whereas those below the y-axis occur less frequently in the sample than would be expected in the proteome. The PSP logo reflects the specificity of the antibody for the S*/T*DXE motif. For more information on motif analysis using PSP, please visit www.phosphosite.org.
The motif logo shows the amino acid distributions around the sites recognized by Immunoafiinity beads from PTMScan® Phospho-Akt Substrate Motif Kit #5561.
The motif logo shows the amino acid distributions around the sites recognized by the Immunoaffinity beads from PTMScan® Phospho-AMPK Substrate Motif (LXRXXS*/T*) Kit #5564.
The motif logo shows the amino acid distributions around the sites recognized by Immunoaffinity beads from PTMScan® Phospho-PKA Substrate Motif (K/RK/RXS*/T*) Kit #5565.
Motif analysis was done using all tyrosine phosphopeptides generated from mouse brain and mouse liver tryptic peptides using Immunoaffinity beads from PTMScan® Phospho-Tyrosine Motif (Y*) (P-Tyr-1000) Kit #8803. The motif logo, drawn from 1178 nonredundant sites, shows the amino acid distributions around the sites recognized by Phospho-Tyrosine (P-Tyr-1000) MultiMab™ Rabbit mAb mix #8954. The motif logo shows that this antibody is a general phospho-tyrosine antibody and does not have amino acid preferences at other positions.
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® 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. An alternate PTMScan® IAP Buffer Plus Detergent #9992 which may reduce nonspecific interactions is available separately. A detailed protocol and Limited Use License allowing the use of the patented PTMScan® method is included with the kit.
Antibody beads supplied in IAP buffer containing 50% glycerol. Store at -20°C. Do not aliquot the antibodies.
PTMScan® Technology employs a proprietary methodology from Cell Signaling Technology for peptide enrichment by immunoprecipitation using a specific bead-conjugated antibody in conjunction with liquid chromatography (LC) tandem mass spectrometry (MS/MS) for quantitative profiling of post-translational modification (PTM) sites in cellular proteins. These include phosphorylation (PhosphoScan®), ubiquitination (UbiScan®), acetylation (AcetylScan®), and methylation (MethylScan®), among others. PTMScan® enables researchers to isolate, identify and quantitate large numbers of post-translationally modified cellular peptides with a high degree of specificity and sensitivity providing a global overview of PTMs in cell and tissue samples without preconceived biases about where these modified sites occur (1). For more information on PTMScan® services, please visit www.cellsignal.com/support.
For more information on our PTMScan® Motif Antibody Kits, please visit www.cellsignal.com/support. Background information specific to each component in the sampler kit can be found on the individual webpages. Phospho-PKA Substrate Motif is described on the webpage for PTMScan® Phospho-PKA Substrate Motif (RRXS*/T*) Kit #5565. Phospho-AMPK Substrate Motif is described on the webpage for PTMScan® Phospho-AMPK Substrate Motif (LXRXXS*/T*) Kit #5564. Phospho-Akt Substrate Motif is described on the webpage for PTMScan® Phospho-Akt Substrate Motif mAb 1 (RXXS*/T*) Kit #5561. Phospho-CK2 Substrate Motif is described on the webpage for PTMScan® Phospho-CK2 Substrate Motif (S*/T*DXE) Kit #12170. Phospho-Tyrosine is described on the webpage for PTMScan® Phospho-Tyrosine Rabbit mAb (P-Tyr-1000) Kit #8803.
AcetylScan is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
MethylScan is a trademark of Cell Signaling Technology, Inc.
MultiMab is a trademark of Cell Signaling Technology, Inc.
PTMScan is a trademark of Cell Signaling Technology, Inc.
UbiScan is a trademark of Cell Signaling Technology, Inc.
Use of Cell Signaling Technology (CST) Motif Antibodies within certain methods (e.g., U.S. Patents No. 7,198,896 and 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 firstname.lastname@example.org.