Anti-mono methyl Arginine 抗体 [5D1] (ab415)

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Key features and details

  • Mouse monoclonal [5D1] to mono methyl Arginine
  • Suitable for: WB, ChIP
  • Reacts with: Species independent
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-mono methyl Arginine antibody [5D1]
    mono methyl Arginine 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [5D1] to mono methyl Arginine

  • 由来種

    Mouse

  • アプリケーション

    適用あり: WB, ChIPmore details

  • 種交差性

    交差種: Species independent

  • 特記事項

    This antibody will be of central importance in analysing the methylation status of chromatin and transcription factors. Since it recognises monomethyl arginine only it can be used in parallel with ab413 (detects dimethyl arginine only) and ab412 (detects both monomethyl and dimethyl arginine) to monitor the exact modification status.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab415 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB Use at an assay dependent dilution.
ChIP Use 5µl for 106 cells.

ターゲット情報

  • 別名

    • Arginine (mono methyl) antibody

画像

  • ChIP - Anti-mono methyl Arginine antibody [5D1] (ab415)
    ChIP - Anti-mono methyl Arginine antibody [5D1] (ab415)This image is courtesy of Sylvain Daujat, Tony Kouzarides lab, Cambridge University, UK

    Sonicated Chromatin prepared from untreated or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure (see protocol) with ab415 to mono methyl Arginine.

    Immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are % of inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 5 µl of ab415 and 4x106 cells were used in each ChIP experiment.

  • ChIP - Anti-mono methyl Arginine antibody [5D1] (ab415)
    ChIP - Anti-mono methyl Arginine antibody [5D1] (ab415)

    We looked at chicken chromatin and then analysed 4 active genes (AG1-4), 2 inactive genes (IG1-2) as well as heterochromatin (Het). A "Yes" indicates that the antibody immunoprecipitated material well above background and should be regarded as a positive result.

     

参考文献

ab415 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab415 は 4 報の論文で使用されています。

  • Cheung P  et al. Single-Cell Chromatin Modification Profiling Reveals Increased Epigenetic Variations with Aging. Cell 173:1385-1397.e14 (2018). PubMed: 29706550
  • Slade DJ  et al. Chemical and biological methods to detect post-translational modifications of arginine. Biopolymers 101:133-43 (2014). PubMed: 23576281
  • Raijmakers R  et al. Methylation of arginine residues interferes with citrullination by peptidylarginine deiminases in vitro. J Mol Biol 367:1118-29 (2007). PubMed: 17303166
  • Cuthbert GL  et al. Histone deimination antagonizes arginine methylation. Cell 118:545-53 (2004). PubMed: 15339660

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Question

samples; HEK293 cells block: 5% milk ab413, ab415: black membrane, protein ladder in white Ab 1/1000 suggestion: use less primary and/or secondary antibody ab109451: lots of bands, Ab 1/1000 suggestion: use less primary antibody (recommended dilution is 1/10,000 to 1/50,000)

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Answer

Thank you for your phone call. I am glad to hear that for ab109451 the protocol tips did work. I am very sorry that unfortunately, for ab413 and ab415, the tips did not improve your results. However, the details provided enable us to closely monitor the quality of our products. I am sorry these 2 products did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued free of charge replacements with the order number xxx for ab107495 and ab107514. You should receive both vials tomorrow. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

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Question

samples; HEK293 cells block: 5% milk ab413, ab415: black membrane, protein ladder in white Ab 1/1000 suggestion: use less primary and/or secondary antibody ab109451: lots of bands, Ab 1/1000 suggestion: use less primary antibody (recommended dilution is 1/10,000 to 1/50,000)

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Answer

Thank you for your phone call. I am sorry to hear that these 3 antibodies are not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. As we discussed over the phone, I would like to offer some suggestions to help optimize the results: For ab413, ab415 I'd suggest using less primary and/or secondary antibody as the membrane shouldn't turn black. This might be an indication of too much antibody on the membrane. For ab109451, most likely using less primary antibody could help with the background as the recommended dilution for the antibody is 1/10,000 to 1/50,000. Should the suggestions not improve the results, please do let me know. In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund. I hope this information is helpful, and I thank you for your cooperation.

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Question

Concerning ab415, ab414, ab413 - says whole antiserum; is it tisue culture supernatant or ascites? Do ab412 and ab413 cross-reat with symmetrical Ng/Na - dimethyl arginine?

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Answer

Thank you for your patience. Ab415, ab414, and ab413 are ascites. It has not been determinded as to whether ab412 and ab413 cross-react with symmetrical Ng/Na - dimethyl arginine.

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