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PTMScan® Control Peptides Mono-Methyl Arginine #40061

    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 Mono-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 Mono-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 Mono-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 Mono-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 Mono-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).

    For Research Use Only. Not For Use In Diagnostic Procedures.
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