Cell Signaling Technology

Product Pathways - Screening Technologies

PTMScan® Phospho-Ser-Pro-Pro Motif [pSPP] Kit #14989

SPP  

REACTIVITY

No. Size Price
14989S 1 Kit ( 10 assays ) 请询价 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
PTMScan® Phospho-Ser-Pro-Pro [pSPP] Immunoaffinity Beads 80 µl Rabbit IgG
PTMScan® IAP Buffer (10X) #9993 600 µl
PTMScan® Limited Use License license

Homology

Species predicted to react based on 100% sequence homology: All Species Expected,

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

Chart

Chart

This chart shows the underlying motif distribution in a PhosphoScan® LC-MS/MS experiment using 750 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-Ser-Pro-Pro Motif [pSPP] Immunoaffinity Beads.

Motif Logo

Motif Logo

The Motif Logo was generated from a PhosphoScan® LC-MS/MS experiment using 750 nonredundant tryptic peptides derived 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-Ser-Pro-Pro Motif [pSPP] Immunoaffinity Beads. The logo demonstrates the relative frequency of amino acids in a given position around the modified site 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.

Background

The MAPK and CDK families of serine/threonine protein kinases play important roles in cell signaling and cell cycle control. These kinases phosphorylate threonine or serine followed by a proline residue (1-6). To facilitate the study and discovery of new MAPK and CDK substrates, Cell Signaling Technology has developed antibodies that bind to phospho-threonine or phospho-serine followed by proline.

The S/TPP motif matches the consensus sequence for targeted phosphorylation by Cdc2 (7,8), ERK1, ERK2 (9), SAPK/JNK, CDK5, and GSK3 (10).

  1. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
  2. Seger, R. and Krebs, E.G. (1995) FASEB J 9, 726-35.
  3. Nurse, P. (2000) Cell 100, 71-8.
  4. Cross, T.G. et al. (2000) Exp Cell Res 256, 34-41.
  5. Yang, C.C. et al. (1998) J Protein Chem 17, 329-35.
  6. Reynolds, C.H. et al. (2000) J Neurochem 74, 1587-95.
  7. Shah, O.J. et al. (2003) J Biol Chem 278, 16433-42.
  8. Songyang, Z. et al. (1994) Curr Biol 4, 973-82.
  9. Gonzalez, F.A. et al. (1991) J Biol Chem 266, 22159-63.
  10. Yang, X. and Gabuzda, D. (1998) J Biol Chem 273, 29879-87.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products


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 ptmscan@cellsignal.com.

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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