Product Pathways - Screening Technologies
PTMScan® Phospho-Tyrosine Mouse mAb (P-Tyr-100) Kit #5636
|5636S||1 Kit ( 5 assays )||email@example.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|
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.
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 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.
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.
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For Research Use Only. Not For Use In Diagnostic Procedures.