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I'm contacting you, since we are having problems with it. As you can see from the attached Western blot image, the antibody seems not to be specific. In particular:
1) the Western Blot was done in TBST-5% milk, by using the antibody o/n at the indicated concentration.
2) we used the following cell lysates:
_ mES (to verify the possible usage in murine cells)
_ NHEK (our human cell line of interest)
_ NHEK shDGCR8 (in which the expression of our protein of interest in constitutively down-regulated).
3) we performed in parallel the Western blotting with another Abcam antibody (ab36865), which according to the shown data sheets on your website could be less specific thanab90579.
As you can see from the image, theab36865 antibody specifically recognized DGCR8 (indicated by the red arrow), while theab90579 did not.
Do you think that there is a problem with the specific lot (GR 34642-6) that we received? And in this case, can you replace it with another one?
Thank you in advice for your willingness and help!
Asked on Sep 25 2012
Thank you for contacting Abcam.
I am sorry about the issues you have been having with ab90579 in western blot. The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on human (but not mouse) samples.
Based on the information that you have provided, I do not think that any protocol tips that I could give would help resolve the issue, therefore I would be happy to replace or refund the antibody for you. Currently we only have the same lot of ab90579 in stock as you originally received, therefore it may be better to either receive a different DGCR8 antibody (possible another vial of ab36865), or any other that you are interested in. Or I could process a refund.
Please let me know how you would you like to continue and if you do wish to receive a replacement the catalogue number of that antibody.
I look forward to your reply and helping to resolve the issue.
Answered on Sep 25 2012