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  3. PTMScan® Control Peptides Phospho-Akt (RXXS*/T*)

PTMScan® Control Peptides Phospho-Akt (RXXS*/T*) #62248

    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 Phospho-Akt (RXXS*/T*) 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 Phospho-Akt (RXXS*/T*) 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 Phospho-Akt (RXXS*/T*) 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 Phospho-Akt (RXXS*/T*) in the LCMS data file.

    Storage

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

    Product Description

    The PTMScan® Control Peptides Phospho-Akt (RXXS*/T*) 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

    Akt plays a central role in mediating critical cellular responses, including cell growth and survival, angiogenesis, and transcriptional regulation (1-3). It is a member of an important class of kinases, referred to as Arg-directed kinases or AGC-family kinases, which includes cAMP-dependent protein kinase (PKA), cGMP-dependent protein kinase (PKG), protein kinase C, Akt, p70 S6 kinase, and RSK. These kinases share a substrate specificity characterized by Arg at position -3 relative to the phosphorylated Ser or Thr (4,5). Akt, p70 S6 kinase, and RSK additionally share specificity for Arg at position -5 (recognition sequence RXRXXS/T) (6). In a recent phosphoproteomic study (7) co-authored by scientists in the Cell Signaling Technology Site Discovery Group, over 300 downstream substrates for AGC-family kinases recognizing the RXRXXS/T motif were identified with PTMScan® Technology using Phospho-Akt substrate antibodies. These Cell Signaling Technology antibodies are powerful tools for investigating the regulation of phosphorylation by Akt and other Arg-directed kinases, as well as for high throughput kinase drug discovery.

    In this assay, PTMScan® (RXXS*/T*) Motif Antibody bead conjugates are used to specifically enrich phosphopeptides containing the RXXS*/T* motif (S* = phospho-serine, T* = phospho-threonine).

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