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68517
PTMScan® Control Peptides Symmetric Di-Methyl Arginine
Proteomic Analysis Products
Cell Extracts

PTMScan® Control Peptides Symmetric Di-Methyl Arginine #68517

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Annotated tandem mass spectrum (MS2) of the PTMScan® Control Peptides Symmetric Di-Methyl Arginine component: GLGR(me2s)GGGGPEGGGFPNGAGPAE[R].
Annotated tandem mass spectrum (MS2) of the PTMScan® Control Peptides Symmetric Di-Methyl Arginine component: GR(me2s)GTPAYPESTFGS[R].
Annotated tandem mass spectrum (MS2) of the PTMScan® Control Peptides Symmetric Di-Methyl Arginine component: SYSR(me2s)GGGDQGYGSG[R].
Peptides included in the PTMScan® Control Peptides Symmetric Di-Methyl Arginine mix.  All peptides are stable-isotope labeled, designated by bracketed R, and contain a di-methyl group designated by parentheses.
Extracted ion chromatograms of PTMScan® Control Peptides Symmetric Di-Methyl Arginine added at supplied concentration (1X at 200 fmol) to human Hep G2 peptides prior to immunoaffinity enrichment using PTMScan® Symmetric Di-Methyl Arginine Motif Kit #13563.  Desalted peptides were analyzed on Q Exactive mass spectrometer and resolved using a 120 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 Symmetric Di-Methyl Arginine 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 Symmetric Di-Methyl Arginine 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 Symmetric Di-Methyl Arginine 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 Symmetric Di-Methyl Arginine 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 Symmetric Di-Methyl Arginine 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

Arginine methylation is a prevalent PTM found on both nuclear and cytoplasmic proteins. Arginine methylated proteins are involved in many different cellular processes, including transcriptional regulation, signal transduction, RNA metabolism, and DNA damage repair (1-3). Arginine methylation is carried out by the arginine N-methyltransferase (PRMT) family of enzymes that catalyze the transfer of a methyl group from S-adenosylmethionine (AdoMet) to a guanidine nitrogen of arginine (4). There are three different types of arginine methylation: asymmetric dimethylarginine (aDMA, omega-NG,NG-dimethylarginine), where two methyl groups are placed on one of the terminal nitrogen atoms of the guanidine group of arginine; symmetric dimethylarginine (sDMA, omega-NG,NG-dimethylarginine), where one methyl group is placed on each of the two terminal guanidine nitrogens of arginine; and monomethylarginine (MMA, omega-NG-methylarginine), where a single methyl group is placed on one of the terminal nitrogen atoms of arginine. Each of these modifications has potentially different functional consequences. Though all PRMT proteins catalyze the formation of MMA, Type I PRMTs (PRMT1, 3, 4, 6, and 8) add an additional methyl group to produce aDMA, while Type II PRMTs (PRMT5 and 7) produce sDMA. Methylated arginine residues often reside in glycine-arginine rich (GAR) protein domains, such as RGG, RG, and RXR repeats (5). However, PRMT4/CARM1 and PRMT5 methylate arginine residues within proline-glycine-methionine rich (PGM) motifs (6). t

In undifferentiated mouse embryonic neural precursors, sDMA histone H4R3 is prevalent, but in later stages of development, both sDMA and aDMA H4R3 modifications are detected in post-mitotic neurons and developing oligodendrocytes. This implies that sDMA modifications may be negative epigenetic regulatory events while aDMA modifications may signal epigenetic activation sites (7).

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 # 68517
Cat. # Size Qty. Price
68517S
100 µl  (10 assays)