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8803
PTMScan® Phospho-Tyrosine Rabbit mAb (P-Tyr-1000) Kit
Proteomic Analysis Products
PTMScan

PTMScan® Phospho-Tyrosine Rabbit mAb (P-Tyr-1000) Kit #8803

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PTMScan® Phospho-Tyrosine Rabbit mAb (P-Tyr-1000) Kit: Image 1

Representative motif analysis of 2362 phospho-tyrosine tryptic peptides generated from a PhosphoScan® experiment of Jurkat cells treated with Calyculin A #9902 and pervanadate using PTMScan® Phospho-Tyrosine Motif (Y*) (P-Tyr-1000) Immunoaffinity Beads. Motif logo was generated using the PhosphoSitePlus® Logo Generator and reflects relative frequency of residues in each position. For more information, please visit PhosphoSitePlus® at www.phosphosite.org.

PTMScan® Phospho-Tyrosine Rabbit mAb (P-Tyr-1000) Kit: Image 2

Motif analysis was done using all tyrosine phosphopeptides generated from a PhosphoScan® experiment of mouse brain and mouse liver tryptic peptides using PTMScan® Phospho-Tyrosine Motif (Y*) (P-Tyr-1000) Immunoaffinity Beads. The Motif Logo, drawn from 1178 nonredundant sites, shows the amino acid distributions around the sites recognized by Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb #8954. The motif logo shows that this antibody is a general phospho-tyrosine antibody and does not have amino acid preferences at other positions.

Product Includes Volume (with Count)
PTMScan® Phospho-Tyrosine Motif (Y*) (P-Tyr-1000) Immunoaffinity Beads 10 x 80 µl
PTMScan® IAP Buffer (10X) 9993 10 x 600 µl
PTMScan® Limited Use License 1 x 1 ml

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 are 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

PhosphoScan® Technology employs phospho-residue (Tyr, Ser, Thr) motif antibodies for phosphopeptide immunoaffinity purification from cell extracts combined with LC tandem MS/MS to identify and quantify changes in phosphorylation levels (1). Tyrosine phosphorylation plays a key role in cellular signaling. Research studies have shown that in cancer, unregulated tyrosine kinase activity can drive malignancy and tumor formation by generating inappropriate proliferation and survival signals (2). Antibodies specific for phospho-tyrosine have been invaluable reagents in these studies (3,4). The Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb #8954, developed by Cell Signaling Technology, provides an exceptionally sensitive tool for studying tyrosine phosphorylation and monitoring tyrosine kinase activity in high throughput drug discovery. PTMScan® (Y*) Motif Antibody bead conjugates are used to specifically enrich phosphopeptides containing Y* motif (Y*= phospho-tyrosine).

  1. Rush, J. et al. (2005) Nat Biotechnol 23, 94-101.
  2. Schlessinger, J. (2000) Cell 103, 211-25
  3. Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65
  4. Ward, S.G. et al. (1992) J Biol Chem 267, 23862-9

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.
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]