The chart shows the relative category distribution of proteins with unmodified or phosphorylated tryptic peptides identified from several combined PTMScan® LC-MS/MS experiments of multiple human samples using PTMScan® Multi-Pathway Immunoaffinity Beads.
The image shows an enlarged schematic representation of the variety of antibodies arrayed on the PTMScan® Multi-Pathway Immunoaffinity protein A bead matrix.
The image shows the hierarchical clustering heat map of mouse liver (left panel), brain (middle panel), and embryo tissue (right panel) from a PTMScan® LC-MS/MS experiment using PTMScan® Multi-Pathway Immunoaffinity Beads.
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® Monoclonal Antibody mix 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. A detailed protocol and Limited Use License allowing the use of the patented PTMScan® method are included with the kit.
Antibody beads supplied in IAP buffer containing 50% glycerol. Store at -20°C. Do not aliquot the antibody.
PTMScan® Technology employs a proprietary methodology from Cell Signaling Technology (CST) 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® Technology 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. For more information on PTMScan® Proteomics Services, please visit www.cellsignal.com/common/content/content.jsp?id=ptmscan-services.
PTMScan® Multi-Pathway Enrichment Kit introduces Cell Signaling Technology's PTMScan® Direct Multi-Pathway proteomics service to the end user in a convenient and easy-to-use kit format. Based on the patented PTMScan® method, the kit includes an array of site-specific antibodies conjugated to protein A beads for peptide immunoaffinity purification. The kit is designed for the screening, discovery, and quantitation of thousands of proteins and phosphorylation sites on hundreds of critical signaling proteins spanning multiple signaling pathways. These include cell cycle and checkpoint control, PI3 Kinase and Akt signaling, MAPK and JNK cascades, T-cell and B-cell receptor signaling, as well as some housekeeping targets. The kit can also be used as the enrichment reagent for customized targeted assays by including heavy isotope-labeled synthetic peptides to a subset of validated targets.
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.
PhosphoScan 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 [email protected] For information regarding commercial licensing terms please contact CST Pharma Services Department at [email protected]