Anti-DNA Ligase IV antibody (ab26039)

The purchase from the university of Maryland on 07/02/06, order number 126743 by Jay Hunter is correct. And in the absence of detection of Artemis and DNA ligase IV using ab18319 and ab26039, I ordered other antibodies for Artemis and DNA-Ligase IV and it worked very well although using the commercially available extraction buffers and blocking buffers. Best Regards

ANSWER:

Thank you for getting back in touch with me with your order details.

I have arranged for a credit note to be raised by our accounts department against order numbers 126743 and 114423 for two vials of ab17988. They will be in touch with your purchasing agent shortly.

With regards the antibodies against Artemis and DNA-Ligase IV (ab18319 and ab26039) I do not have details of your order previously. If you are referring to other commercially available antibodies against these targets then I must stress that the behaviour of one antibody is not necessary transferable to another. I would still like to recommend that these antibodies are applied as detailed using RIPA buffer extraction of the human cells that you have been using; verify the transfer of your protein by ponceau staining and use 3% BSA/TBS (0.1% tween 20) as a blocking agent.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Dear abcam! I hope it works this time.

ANSWER:

Thank you for your enquiry.

In Charlotte's absence I am dealing with her enquiries. I am sorry to hear that you have been having difficulties with these antibodies.

I have been reading through your technical questionaires and I appreciate the detail that you have provided me with. I am unaware of the commercial extraction agents that you have been employing. However, I am certainly concerned about the extraneous bands that you have been observing when using our Werner's syndrome helicase WRN antibody (ab17988). I am certainly prepared to provide you with credit to the value of one vial of each of these antibodies provided that they have been purchased within the past 90 days.

With respect to your observed absence of detection of Artemis and DNA Ligase IV using ab18319 and ab26039 respectively I would like to suggest that you use a RIPA buffer extraction of the human cells that you have been using; verify the transfer of your protein by ponceau staining and use 3% BSA/TBS (0.1% tween 20) as a blocking agent. My concern is that the commercially available extraction buffers and blocking buffers that you have been using are perhaps not the best agents for the detection of these proteins. It would therefore certainly help to prepare fresh buffers prior to an immunoblotting experiment.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.