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

Submit a review Q&A (3)References (4)

Overview

  • Product name

    Anti-mono methyl Arginine antibody [5D1]
    See all mono methyl Arginine primary antibodies

  • Description

    Mouse monoclonal [5D1] to mono methyl Arginine

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, ChIPmore details

  • Species reactivity

    Reacts with: Species independent

  • General notes

    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.

Properties

Applications

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.

Application Abreviews Notes
WB Use at an assay dependent dilution.
ChIP Use 5µl for 106 cells.

Target

  • Alternative names

    • Arginine (mono methyl) antibody

Images

  • 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.

     

References

This product has been referenced in:

  • Cheung P  et al. Single-Cell Chromatin Modification Profiling Reveals Increased Epigenetic Variations with Aging. Cell 173:1385-1397.e14 (2018). Read more (PubMed: 29706550) »
  • Slade DJ  et al. Chemical and biological methods to detect post-translational modifications of arginine. Biopolymers 101:133-43 (2014). Read more (PubMed: 23576281) »
See all 4 Publications for this product

Publishing research using ab415? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

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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