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5566
PTMScan® Phospho-ST*P Motif (ST*P) XP® Rabbit mAb Kit
Proteomic Analysis Products
PTMScan

PTMScan® Phospho-ST*P Motif (ST*P) XP® Rabbit mAb Kit #5566

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PTMScan® Phospho-ST*P Motif (ST*P) XP® Rabbit mAb Kit: Image 1
The Motif Logo shows the amino acid distributions around the sites recognized by the antibody.
PTMScan® Phospho-ST*P Motif (ST*P) XP® Rabbit mAb Kit: Image 2
Chart showing the proportion of underlying sequence motifs found in a PTMScan® study using a PLK binding motif antibody and OrbiTrap MS analysis. Analysis of peptides from HeLa cells, untreated and nocodazole-treated, gave 452 non-redundant sites containing a PLK binding and related motifs. The primary motif is highlighted in white 98% of these sites fit the ST*P motif; only 2% of the sites are from other phospho-Ser/Thr containing peptides that do not have the motif, which shows that this antibody is very specific for the ST*P motif.
Product Includes Volume (with Count)
PTMScan® Phospho-PLK Binding Motif (ST*P) Immunoaffinity Beads 10 x 80 µl
PTMScan® IAP Buffer (10X) 9993 10 x 600 µl
PTMScan® Limited Use License 1 x 1 ea

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® 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 is included with the kit.

Storage

Antibody beads supplied in IAP buffer containing 50% glycerol. Store at -20°C. Do not aliquot the antibody.

Product Description

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 (1). For more information on PTMScan® Proteomics Services, please visit www.cellsignal.com/common/content/content.jsp?id=ptmscan-services.

Background

The ST*P motif is implicated in signaling by polo-like kinases (PLKs). These Ser/Thr protein kinases play essential roles during the cell cycle. At least four PLKs exist in mammalian cells: PLK1, PLK2, PLK3, and PLK4 (2). PLKs have a highly conserved N-terminal kinase domain and a relatively divergent C-terminal domain called the Polo-box domain (PBD). Of the four PLKs, PLK1 is the best characterized (3). PLK1 functions as a key regulator of mitotic events by phosphorylating substrate proteins on centrosomes, kinetochores, the mitotic spindle, and the midbody, and is crucial for proper progression through multiple stages of mitosis (4-6).
The PBDs of PLK1 function as a phospho-Ser/Thr-binding module, optimally recognizing the sequence motif S[S*/T*][P/X] (PLK1 binding motif) (6). Binding of phosphopeptides containing S[S*/T*][P/X] motif by the PBD in PLK1 relieves its inhibitory function on kinase activity (7). S[S*/T*]P peptides are phosphorylated by proline-directed kinases such as cyclin dependent kinases (CDKs) (6). These findings imply that priming phosphorylations on substrates or docking proteins by other mitotic kinases such as CDKs may target PLK1 to its substrates and simultaneously activate its kinase domain. The ST*P Motif (ST*P) XP® Rabbit mAb contained in this kit, which binds to ST*P, was developed by Cell Signaling Technology and is a useful tool to study PLK1 binding proteins and PLK1 substrates.
In this assay, PTMScan® (ST*P) Motif Antibody bead conjugates are used to specifically enrich phosphopeptides containing the ST*P motif (T* = phospho-threonine).
  1. Rush, J. et al. (2005) Nat Biotechnol 23, 94-101.
  2. Nigg, E.A. (1998) Curr Opin Cell Biol 10, 776-83.
  3. Donaldson, M.M. et al. (2001) J Cell Sci 114, 2357-8.
  4. Barr, F.A. et al. (2004) Nat Rev Mol Cell Biol 5, 429-40.
  5. Kishi, K. et al. (2009) Mol Cell Biol 29, 3134-50.
  6. Elia, A.E. et al. (2003) Science 299, 1228-31.
  7. Elia, A.E. et al. (2003) Cell 115, 83-95.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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.
PTMScan is a trademark of Cell Signaling Technology, Inc.
UbiScan is a trademark of Cell Signaling Technology, Inc.
XP is a registered 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]