Product Pathways - Screening Technologies
PTMScan® Phospho-Tyrosine Mouse mAb (P-Tyr-100) Kit #5636
| No. | Size | Ordering Information | |
|---|---|---|---|
| 5636S | 1 Kit ( 5 assays ) |  |
ptmscan@cellsignal.com |
| Products Included | Quantity | Cap Color |
|---|---|---|
| PTMScan® (Y*) Motif Antibody Bead Conjugate | 5 x 80 µl | |
| PTMScan® IAP Buffer (10X) | 5 x 0.6 ml | White |
| PTMScan® Limited Use License |
PTMScan® Services
View sample data from a PTMScan® study using the Motif Antibody in this kit, provided in a PTMScan® Service report.
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® 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 is included with the kit.
Description
PTMScan® Technology employs a proprietary methodology from Cell Signaling Technology 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® 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® services, please visit www.cellsignal.com/services/index.html.
Motif Logo
Motif analysis was done using all tyrosine phosphopeptides generated from studies in the published report by Rikova, K.A., et al. (2007) Cell 131, 1190-1203. The Motif Logo, drawn from 4160 non-redundant sites, shows the amino acid distributions around the sites recognized by Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411. The motif analysis shows that this antibody is a general phospho-tyrosine antibody and does not have amino acid preferences at other positions.
Background
Tyrosine phosphorylation plays a key role in cellular signaling (2). In cancer, unregulated tyrosine kinase activity can drive malignancy and tumor formation by generating inappropriate proliferation and survival signals (3). Antibodies specific for phospho-tyrosine (4,5) have been invaluable reagents in these studies. The Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411, developed by Cell Signaling Technology, provides an exceptionally sensitive tools for studying tyrosine phosphorylation and monitoring tyrosine kinase activity in high throughput drug discovery.In this assay, PTMScan® (Y*) Motif Antibody bead conjugates are used to specifically enrich phosphopeptides containing *Y motif (Y* = phospho-tyrosine).
- Rush, J. et al. (2005) Nat Biotechnol 23, 94-101.
- Schlessinger, J. (2000) Cell 103, 211-25.
- Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.
- Ward, S.G. et al. (1992) J Biol Chem 267, 23862-9.
- Glenney, J.R. et al. (1988) J Immunol Methods 109, 277-85.
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!
For Research Use Only. Not For Use In Diagnostic Procedures.
