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  3. PTMScan® Control Peptides Ubiquitin/SUMO

PTMScan® Control Peptides Ubiquitin/SUMO #75964

    Product Information

    Product Usage Information

    Use with Cell Signaling Technology’s PTMScan® kit protocol from the Immunoaffinity Purification (IAP) step. Because the optimal amount of PTMScan® Control Peptides Ubiquitin/SUMO for each user’s experiments will depend on unique factors such as mass spectrometer sensitivity, users may dilute these control peptides as needed.

    1. Aliquot PTMScan® Control Peptides Ubiquitin/SUMO for storage as single-use units at -20°C or proceed to immediate usage.
    2. Resuspend sample peptides in the appropriate buffer and volume, e.g., 1.4 mL of PTMScan® IAP Buffer (1X).
    3. Clear sample peptides by centrifugation.
    4. Transfer clarified sample peptides to tubes containing IAP beads.
    5. Add 10 µL of PTMScan® Control Peptides Ubiquitin/SUMO to IAP beads and sample peptides and mix well.
    6. Continue with PTMScan® or PTMScan® HS workflows at the 2-hour incubation step.
    7. Detect PTMScan® Control Peptides Ubiquitin/SUMO in the LCMS data file.

    Storage

    This product is stable for 12 months when stored at -20°C. Aliquot to avoid multiple freeze/thaw cycles.

    Product Description

    The PTMScan® Control Peptides Ubiquitin/SUMO enable quality control of immunoaffinity enrichment performance using PTMScan® or PTMScan® HS workflows. These synthetic peptides contain a specific post-translational modification (PTM) that can be enriched by the associated PTMScan® or PTMScan® HS immunoaffinity purification (IAP) beads, as well as a stable heavy isotope that can be distinguished from endogenous peptides by the mass spectrometer.

    Background

    Ubiquitin is a conserved polypeptide unit that plays an important role in the ubiquitin-proteasome pathway. Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the activation component E1; the activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (1-3). The ubiquitin-proteasome pathway has been implicated in a wide range of normal biological processes and in disease-related abnormalities. Several proteins such as IκB, p53, cdc25A, and Bcl-2 have been shown to be targets for the ubiquitin-proteasome process as part of regulation of cell cycle progression, differentiation, cell stress response, and apoptosis (4-7).

    Small ubiquitin-related modifier 1, 2, and 3 (SUMO-1, -2, and -3) are members of the ubiquitin-like protein family (8). The covalent attachment of the SUMO-1, -2, or -3 (SUMOylation) to target proteins is analogous to ubiquitination.

    Ubiquitin and the individual SUMO family members are all targeted to different proteins with diverse biological functions. Ubiquitin predominantly regulates degradation of its target (8). In contrast, SUMO-1 is conjugated to RanGAP, PML, p53, and IκB-α, regulates nuclear trafficking, forms subnuclear structures, and regulates transcriptional activity and protein stability (9-13). SUMO-2/-3 forms poly-(SUMO) chains, is conjugated to topoisomerase II and APP, regulates chromosomal segregation and cellular responses to environmental stress, and plays a role in the progression of Alzheimer's disease (14-17).

    1. Ciechanover, A. (1998) EMBO J 17, 7151-60.
    2. Hochstrasser, M. (2000) Nat Cell Biol 2, E153-7.
    3. Hochstrasser, M. (2000) Science 289, 563-4.
    4. Bernardi, R. et al. (2000) Oncogene 19, 2447-54.
    5. Aberle, H. et al. (1997) EMBO J 16, 3797-804.
    6. Salomoni, P. and Pandolfi, P.P. (2002) Nat Cell Biol 4, E152-3.
    7. Jesenberger, V. and Jentsch, S. (2002) Nat Rev Mol Cell Biol 3, 112-21.
    8. Schwartz, D.C. and Hochstrasser, M. (2003) Trends Biochem Sci 28, 321-8.
    9. Matunis, M.J. et al. (1996) J Cell Biol 135, 1457-70.
    10. Duprez, E. et al. (1999) J Cell Sci 112 (Pt 3), 381-93.
    11. Gostissa, M. et al. (1999) EMBO J 18, 6462-71.
    12. Rodriguez, M.S. et al. (1999) EMBO J 18, 6455-61.
    13. Desterro, J.M. et al. (1998) Mol Cell 2, 233-9.
    14. Tatham, M.H. et al. (2001) J Biol Chem 276, 35368-74.
    15. Azuma, Y. et al. (2003) J Cell Biol 163, 477-87.
    16. Li, Y. et al. (2003) Proc Natl Acad Sci U S A 100, 259-64.
    17. Saitoh, H. and Hinchey, J. (2000) J Biol Chem 275, 6252-8.

    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 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 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.
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    PTMScan is a registered trademark of Cell Signaling Technology, Inc.
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