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7900
PhosphoScan® Kit (P-Tyr-100)
Cellular Assay Kits
Assay Kit

PhosphoScan® Kit (P-Tyr-100) #7900

Citations (12)

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PhosphoScan® Kit (P-Tyr-100): Image 1

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Product Description

PhosphoScan® Kit (P-Tyr-100) from Cell Signaling Technology (CST) allows for the purification and identification of tyrosine phosphorylation sites in cellular proteins when coupled with LC tandem mass spectrometry using CST's patented PhosphoScan® technology. The assay is based on the specific enrichment of phosphotyrosine-containing peptides using an antibody against phosphotyrosine. Cells are lysed in a urea-containing buffer, and cellular proteins are digested by protease and fractionated by reversed-phase solid-phase extraction. Peptides are then subjected to immunoaffinity purification using Phosphotyrosine Mouse mAb (P-Tyr-100) #9411 coupled to protein G agarose beads*. Overnight incubation ensures high-affinity binding of phosphotyrosine-containing peptides to P-Tyr-100 beads. Unbound peptides are removed through washing, and phosphotyrosine-containing peptides are eluted with dilute acid. Reversed-phase chromatography is performed on microtips to separate phosphopeptides from antibody and to concentrate them for LC tandem mass spectrometry. The kit also contains control peptides from 2x108 Jurkat cells that were treated with pervandate and calyculin A. These control peptides allow the investigator to determine the efficiency of the immunoaffinity purification (IAP) step and the LC-MS/MS analysis (1 assay). *Product is specific to this kit.

Background

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. For more information on PTMScan® services, please visit www.cellsignal.com/services/index.html

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). Current phosphoproteomic approaches generally reveal only small numbers of tyrosine phosphorylation sites, in keeping with the low level of phosphotyrosine relative to phosphoserine and phosphothreonine residues (4). PhosphoScan® Kit, based on CST's published method (1,5), isolates large numbers of cellular phosphotyrosine-containing peptides. This allows the investigator to obtain a global overview of tyrosine phosphorylation in the cell, covering many classes of proteins without preconceived biases about where tyrosine phosphorylation sites will be found.

  1. Schlessinger, J. (2000) Cell 103, 211-25.
  2. Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.
  3. Mann, M. et al. (2002) Trends Biotechnol 20, 261-8.
  4. Rush, J. et al. (2003) U.S. Patent Publication , 20030044848.

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.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PhosphoScan is a trademark of Cell Signaling Technology, Inc.