Anti-dimethyl Arginine 抗体 [21C7] (ab413)

Thank you for your enquiry.

I can confirm that ab412 mono and dimethyl Arginine antibody [7E6] cross reacts with both symetric and assymetric methyl arginine. However, the ab413dimethyl Arginineantibodyis specific to ADMA only.

https://www.abcam.com/index.html?datasheet=413 (or use the following: https://www.abcam.com/index.html?datasheet=413).

I hope this will be helpful to you. If you have any further questions, please do not hesitate to let me know.

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.

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.

Thank you for your enquiry. Thank you also for completing our questionnaire so comprehensively. I am sorry that you have been having difficulties with our antisera against methyl arginine. I have examined your questionnaire and it seems as if you are following a protocol not dissimilar to Abcam's recommended protocol. You mentioned that your target protein is methylated. I have discovered that ICP27 is methylated at Arginine residues. I would have mentioned that your immunoprecipitation by H1119 may be obscured by the methylated form of the protein, resulting in the preferential precipitation of the unmethylated form. However, this does not explain the fact that you are NOT obtaining a result following western blotting using a crude HeLa extract. Given a crude extract the very least that I would expect to see would be the arginine methylated histones H3 and H4. In fact this is what I would recommend you use as positive control to guarantee that your abcam antisera are working and detecting methyl-arginine. Preparing a high concentrated histone extract can be done on HeLa cells. We have a very easy protocol at the link below. This will not affect your protocol given that you are using a 12.5% gel. Histones are between 10 and 17KDa. https://www.abcam.com/index.html?pageconfig=view_protocol&pid=316 You mention that you did have some success using a 1:250 dilution. Please can you e-mail me these scanned as JPeGs. I would also like to know the mass of protein you are loading onto the gel during the whole cell extract western blot and the relative amount you are loading following IP. I would recommend a starting amount of 20ug for the crude extract. Following some literature searching I would also like to know whether your ICP27 construct that you are injecting contains an RGG box as I have learnt that the methylation of ICP27 is dependent on its presence. I look forward to your reply and hope that we can resolve this issue quickly.

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.

Thank you for your enquiry and your interest in our products. We are sorry to inform you that there is no protein concentration for polyclonal antibodies that are not affinity purified. This antibody is provided as whole serum and the IgG concentration has not been determined.

This antibody has not been tested in ELISA. All the latest information that we have about this antibody is found on the datasheet.

The epitope is di-methyl arginine independent of the surrounding sequence and has been used on numerous different diMeArg peptides.

Sorry, but the immunogen of this antibody is confidential. All we can say is that it contained methylated arginine (the antibody is inhibited by the modified amino acid).