Overview

  • Product name
    Anti-5-Hydroxymethyluridine antibody
  • Description
    Goat polyclonal to 5-Hydroxymethyluridine
  • Host species
    Goat
  • Tested applications
    Suitable for: IP, ELISAmore details
  • Immunogen

    5-Hydroxymethyluridine coupled to a carrier protein.

Properties

Applications

Our Abpromise guarantee covers the use of ab19735 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
IP Use at an assay dependent concentration. PubMed: 22514282
ELISA 1/25000.

Target

  • Relevance
    5 Hydroxymethyluridine is formed from thymidine during the oxidation of DNA by ionizing radiation or reactive oxygen species (ROS) that are formed from inflammatory response. This causes genetic mutations, altered gene expression and chromosomal instability that has generally been implicated with aging, cancers and autoimmune disease.
  • Alternative names
    • 5 HmdU antibody
    • 5 Hydroxymethyl 2’ deoxyuridine antibody

References

This product has been referenced in:
  • Cliffe LJ  et al. JBP1 and 2 are Fe2+/2-OG dependent dioxygenases regulating the hydroxylation of thymidine residues in trypanosome DNA. J Biol Chem : (2012). IP . Read more (PubMed: 22514282) »
See 1 Publication for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number XXXXXX. 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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Answer

Thank you for contacting us.


While Dot Blot does indeed have a lot of similarity to a non-reduced/denatured western blot or with an indirect ELISA, we do consider it to be a separate application from either of these. We do however recognize that many products have not been tested for use in this application and as such using a product which is guaranteed for use in one of these is probably the best bet when searching for antibodies.

If you could provide me with the details of your protocol and with any troubleshooting steps and images which you may have, I would be very happy to review these and see if I might be able to provide suggestions to improve the performance of these products.While these are not guaranteed for use in Dot Blot, should we not be able to provide help with these I am sure we might be able to find a way to come to a result which would be satisfactory to both Abcam and you.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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https://www.abcam.com/abreviews

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Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab19735. 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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Answer

Thanks for your reply. Actually you have just been corresponding with me regarding this issue. My name is Jeremy and my signature can be found at the bottom of these emails. While you have been sending emails to our general Scientific Support inbox at us.technical@abcam.com, our system directs emails internally to the individual email account of the Scientific Support Specialist that sent you the initial email. I do recall we discussed the troubles you were having with your western blot experiment. Perhaps there is some confusion here and I'm sorry if I am mistaken but I believe the antibody used in that western blot was ab93155. This product number corresponds to the only antibody purchased on order 964150, the order number you mentioned in your most recent email. As you suggest, perhaps it would be more effiecient to discuss this over the phone. Please feel free to call me directly at 617-577-4244 when you have a chance. If there is a problem with an antibody then I would be happy to send you a free replacement. I look forward to hearing from you.

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Answer

Thanks for your reply. I don't believe we discussed the details of your ELISA experiment using ab19735. For our records, please provide a few details regarding the experiment including the sample type and preparation methods and a brief description of the protocol used as well as the antibody-related problems. Thanks in advance for the additional information!  

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Answer

Thank you for your enquiry. We certainly do guarantee our antibodies to work as noted on our online product datasheets. At this time ab19735 is described as having been tested in ELISA. So if the vial of ab19735 is not working in ELISA then I would be happy to send you a replacement of ab93155 as you have requested. In your reply, please include the purchase order number or Abcam order reference number associated with the order of ab19735 so I can process your request.

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Answer

Thank you for your enquiry. The immunogen used for Goat anti 5-Hydroxymethyluridine, antibody ab19735, was 5-HmUrd coupled to BSA. The 5-HbUrd immunogen was prepared as follows: The 5-HmUrd was dissolved in sodium periodate and reacted for 20 minutes before adding ethylene glycol and bovine serum albumin. After further incubation for 45 minutes, sodium borohydride was added and incubated for 18 hours followed by dialysis against Tris-buffered saline. The final conjugate is 5-HmUrd-BSA. The laboratory that generated the antibody has not tested it in dot blot against oligos on nitrocellulose, but they have the following thoughts. Because genomic DNA is large, part of the DNA may be absorbed to the nitrocellulose (NC) with part extending outward to allow the antibody access to the DNA. Oligos are much smaller and when absorbed to the NC paper they may lay flat against it and do not allow the antibody access to the 5-HmUrd. If this is true, you may want to try incubation of both the oligo and antibody in a test tube and allow them to bind before absorbing onto the NC. The NC can then be incubated with the secondary antibody, followed by processing as usual. I hope this information helps. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us.

The product will cross react as follows

5-Hydroxy-2-deoxyuridine (5-HmdUrd) 100%

5-Hydroxymethyluridine (5-HmUrd) 100%

Thymidine (T) 60%

Hydroxymethyluracil (HmU) 43%

Deoxyuridine (dUrd) 36%

Uridine (Urd) 32%

Uracil (U) 0%

The antibody will detect DNA containing hydroxymethyluracil only. It wioll bind to hydroxymethyluracil irrespective ofr single stranded or double stranded DNA.

You can use this antibody with any normal ELISA experiment e.g. with HRP or AP conjugated secondary antibodies. However we tested this antibody in peptide ELISA with the peptide used as immunogen.

We have not tested this antibody in dot blot and so I am sorry we are unable to confirm it. If there are evidence that this product was used in dot blot, please feel free to use it.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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