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PTMScan® Mono-Methyl Arginine Motif [mme-RG] Kit
Proteomic Analysis Products
PTMScan

PTMScan® Mono-Methyl Arginine Motif [mme-RG] Kit #12235

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PTMScan® Mono-Methyl Arginine Motif [mme-RG] Kit: Image 1

The pie chart shows the relative category distribution of proteins with mono-methylated arginine identified from peptides generated from a MethylScan® LC-MS/MS experiment of HCT 116 cells using PTMScan® Mono-Methyl Arginine (mme-RG) Immunoaffinity Beads.

PTMScan® Mono-Methyl Arginine Motif [mme-RG] Kit: Image 2

The Motif Logo was generated from a MethylScan® LC-MS/MS experiment using 722 nonredundant tryptic peptides derived from human HCT 116 cells immunoprecipitated with PTMScan® Mono-Methyl Arginine motif [mme-RG] Immunoaffinity Beads. The logo represents the relative frequency of amino acids in each position surrounding the central methylated arginine residue. Glycine residues are enriched, especially at the +1 position, in the context of mono-methylated arginine when compared to the overall expected frequency in the human proteome. Of the total methylated arginine peptides, 68% contain the [mme-RG] motif.

Product Includes Volume (with Count)
PTMScan® Mono-Methyl Arginine Motif [mme-RG] Immunoaffinity Beads 10 x 80 µl
PTMScan® IAP Buffer (10X) 9993 10 x 600 µl
PTMScan® Limited Use License 1 x 1 ml

Product Usage Information

Cells are lysed in a urea-containing buffer, cellular proteins are digested by proteases, and the resulting peptides are purified by reversed-phase solid-phase extraction. Peptides are then subjected to immunoaffinity purification using a PTMScan® Motif Antibody conjugated to protein A agarose beads. Unbound peptides are removed through washing, and the captured PTM-containing peptides are eluted with dilute acid. Reversed-phase purification is performed on microtips to desalt and separate peptides from antibody prior to concentrating the enriched peptides for LC-MS/MS analysis. CST recommends the use of PTMScan® IAP Buffer #9993 included in the kit. A detailed protocol and Limited Use License allowing the use of the patented PTMScan® method are included with the kit.

Storage

Antibody beads supplied in IAP buffer containing 50% glycerol. Store at -20°C. Do not aliquot the antibody.

Product Description

PTMScan® Technology employs a proprietary methodology from Cell Signaling Technology (CST) for peptide enrichment by immunoprecipitation using a specific bead-conjugated antibody in conjunction with liquid chromatography (LC) tandem mass spectrometry (MS/MS) for quantitative profiling of post-translational modification (PTM) sites in cellular proteins. These include phosphorylation (PhosphoScan®), ubiquitination (UbiScan®), acetylation (AcetylScan®), and methylation (MethylScan®), among others. PTMScan® Technology enables researchers to isolate, identify, and quantitate large numbers of post-translationally modified cellular peptides with a high degree of specificity and sensitivity, providing a global overview of PTMs in cell and tissue samples without preconceived biases about where these modified sites occur (1). For more information on PTMScan® Proteomics Services, please visit www.cellsignal.com/common/content/content.jsp?id=ptmscan-services.

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,N’G-dimethylarginine), where one methyl group is placed on each of the two terminal guanidine nitrogens of arginine; and monomethylarginine (MMA, omega-NG-dimethylarginine), 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, and 6) 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).

  1. Bedford, M.T. and Richard, S. (2005) Mol Cell 18, 263-72.
  2. Pahlich, S. et al. (2006) Biochim Biophys Acta 1764, 1890-903.
  3. Bedford, M.T. and Clarke, S.G. (2009) Mol Cell 33, 1-13.
  4. McBride, A.E. and Silver, P.A. (2001) Cell 106, 5-8.
  5. Gary, J.D. and Clarke, S. (1998) Prog Nucleic Acid Res Mol Biol 61, 65-131.
  6. Cheng, D. et al. (2007) Mol Cell 25, 71-83.

使用に関する制限

法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

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For Research Use Only. Not For Use In Diagnostic Procedures.
AcetylScan is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
MethylScan is a trademark of Cell Signaling Technology, Inc.
PTMScan is a trademark of Cell Signaling Technology, Inc.
UbiScan is a trademark of Cell Signaling Technology, Inc.
U.S. Pat. No. 9,085,609
Use of Cell Signaling Technology (CST) Motif Antibodies within certain methods (e.g., U.S. Patents No. 7,198,896 and 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at [email protected] For information regarding commercial licensing terms please contact CST Pharma Services Department at [email protected]