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

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PTMScan® Control Peptides Phospho-Akt (RXXS*/T*): Image 1
Annotated tandem mass spectrum (MS2) of the PTMScan® Control Peptides Phospho-Akt (RXXS*/T*) component: RRPT(p)LGVQLDD[K].
PTMScan® Control Peptides Phospho-Akt (RXXS*/T*): Image 2
Annotated tandem mass spectrum (MS2) of the PTMScan® Control Peptides Phospho-Akt (RXXS*/T*) component: RYPS(p)SISSSPQ[K].
PTMScan® Control Peptides Phospho-Akt (RXXS*/T*): Image 3
Annotated tandem mass spectrum (MS2) of the PTMScan® Control Peptides Phospho-Akt (RXXS*/T*) component: VRYDS(p)SNQV[K].
PTMScan® Control Peptides Phospho-Akt (RXXS*/T*): Image 4
Peptides included in the PTMScan® Control Peptides Phospho-Akt (RXXS*/T*) mix.  All peptides are stable-isotope labeled, designated by bracketed K, and contain a phosphate group designated by parentheses. 
PTMScan® Control Peptides Phospho-Akt (RXXS*/T*): Image 5
Extracted ion chromatograms of PTMScan® Control Peptides Phospho-Akt (RXXS*/T*) added at supplied concentration (1X at 200 fmol) to mouse embryo peptides prior to immunoaffinity enrichment using PTMScan® Phospho-Akt Substrate Motif mAb 1 (RXXS*/T*) Kit #5561. Desalted peptides were analyzed on Q Exactive mass spectrometer and resolved using a 90 min reversed phase gradient from 7.5% to 32% acetonitrile on a C18 column. The peak corresponding to the specific Control Peptide is marked with retention time and observed precursor mass, with peak height reported as the normalized level (NL) for each row per panel.

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. 
CAS 75-05-8

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).
  1. Marte, B.M. and Downward, J. (1997) Trends Biochem Sci 22, 355-8.
  2. Jiang, B.H. et al. (2000) Proc Natl Acad Sci U S A 97, 1749-53.
  3. Scheid, M.P. and Woodgett, J.R. (2000) Curr Biol 10, R191-4.
  4. Montminy, M. (1997) Annu Rev Biochem 66, 807-22.
  5. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
  6. Manning, B.D. and Cantley, L.C. (2007) Cell 129, 1261-74.
  7. Moritz, A. et al. (2010) Sci Signal 3, ra64.

Limited Uses

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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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To Purchase # 62248
Cat. # Size Qty. Price
62248S
100 µl  (10 assays)
N/A

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