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We have been struggling western blot for MeCP2 with the one below.
Although we see bands in our blot usingab2828 below. Bands seem to be non-specific. Could you help with us?
Asked on May 11 2012
Thank you for your enquiry regarding a2828 and ab992 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with these two antibodies.
As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.
a2828: Anti-MeCP2 antibody - ChIP Grade
This item was purchased almost 2 years ago (Order No: 70326, 10 Jun 2010) and I regret to inform you that the Abpromise for this antibody has expired.
After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:
1) MeCP2 is localised to the nuclei so try to prepare nuclear lysate instead of whole cell lysate.
2) It is worth testing different dilutions as well (1/700 or 1/500) to see if the intensity of the signal is getting stronger or not
3) Not sure what cell line was used, this information seems to be missing in the e-mail. It would be worth running cell lysate from HeLa, AtT20 or U2OS.
ab992: Anti-Rad21 antibody - ChIP Grade
The protocol looks fine to me and since this antibody was purchased on 31Oct 2011 our Abpromise will certainly applies. Here are a few comments and tips I could make which may help improve the signal.
1) Try to increase the final concentration of this primary antibody on the datasheet, it is recommend using 2 µg/ml.
2) Rad21 is localised to the nuclei so try to prepare nuclear lysate instead of whole cell lysate.
I hope this will be useful for you. Should you still have any problem with this ab992 after following these suggestions, then please do not hesitate to contact our Technical Department again our Abpromise will apply for ab992.
Answered on May 11 2012