PTMScan^® Phospho-Ser/Thr Motif [pS/T] Kit #25081

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. A detailed protocol and Limited Use License allowing the use of the patented PTMScan^® method are included with the kit.

Storage:

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 https://www.cellsignal.com/services/index.html.

As an integral part of the machinery of cellular function, proteins undergo regulation by a variety of post-translational modifications. One of the most prevalent and widely studied PTMs is Serine/Threonine phosphorylation. A few prominent kinases targeting a handful of substrate consensus motifs account for a majority of the tens of thousands of known and predicted sites on more than 13,000 human proteins (1-3). Cell Signaling Technology has developed phospho-Ser/Thr motif antibodies for proteomic profiling of kinase substrates. These include, among others, substrates of Akt, AMPK, MAPK, CDK, PKA, PKC, and CKII.

1. Rao, R.S. and Møller, I.M. (2012) Biochim Biophys Acta 1824, 405-12.
2. Goel, R. et al. (2012) Mol Biosyst 8, 453-63.
3. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.

• Data sheet (179KB)
• MSDS PDF