Product Pathways - Screening Technologies
PTMScan® Phospho-MAPK Substrate Motif [PXpTP] Kit #14990
|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|PTMScan®Phospho-MAPK Substrate Motif [PXpTP] Immunoaffinity Beads||80 µl||Rabbit IgG|
|PTMScan® IAP Buffer (10X) #9993||600 µl|
|PTMScan® Limited Use License||license|
Species predicted to react based on 100% sequence homology: All Species Expected,
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.
This chart shows the distribution of underlying phospho-peptide motifs in a PhosphoScan® LC-MS/MS experiment using 754 nonredundant tryptic peptides generated from mitotic HeLa cells. HeLa cells were treated with thymidine, (2 mM, 18 hr) followed by Nocodazole #2190 (60 nM, 24 hr) and immunoprecipitated using PTMScan® Phospho-MAPK Substrate Motif [PXpTP] Immunoaffinity Beads.
The Motif Logo was generated from a PhosphoScan® LC-MS/MS experiment using 754 nonredundant tryptic peptides derived from mitotic HeLa cells that were treated with thymidine (2 mM, 18 hr) followed by Nocodazole #2190 (60 nM, 24 hr) and immunoprecipitated using PTMScan® Phospho-MAPK Substrate Motif [PXpTP] Immunoaffinity Beads. The Motif Logo demonstrates the relative abundance of amino acids in a given position within this data set.
Directions for Use
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. 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 are included with the kit.
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 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. Cell Signaling Technology has developed antibodies that bind to phospho-threonine followed by proline, motifs PXS*/T*P and/or S*PXR/K, for use in the study and discovery of new MAPK and CDK substrates (4,5).
- Cross, T.G. et al. (2000) Exp Cell Res 256, 34-41.
- Reynolds, C.H. et al. (2000) J Neurochem 74, 1587-95.
- Seger, R. and Krebs, E.G. (1995) FASEB J 9, 726-35.
- Holmes, J.K. and Solomon, M.J. (1996) J Biol Chem 271, 25240-6.
- Songyang, Z. et al. (1996) Mol Cell Biol 16, 6486-93.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
For Research Use Only. Not For Use In Diagnostic Procedures.
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.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PTMScan is a trademark of Cell Signaling Technology, Inc.
AcetylScan is a trademark of Cell Signaling Technology, Inc.
PhosphoScan is a trademark of Cell Signaling Technology, Inc.
UbiScan is a trademark of Cell Signaling Technology, Inc.
MethylScan is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
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