Description
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 ® 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.
Storage
Antibody beads supplied in IAP buffer containing 50% glycerol. Store at -20°C. Do not aliquot the antibody.
Protocol
Available protocols: PTMScan
Background
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.
