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Asked on Mar 28 2012
Thank you for sending your protocol and the image of your blot. You have already tried most of the protocol modifications I would recommend in this case. The only other suggestion I would make is that it might help to use 5% milk for both the blocking buffer and antibody diluents. You may also find that the background signal is reduced by increasing the blocking time to 2-3 hours or overnight at 4C.
After reviewing the image you sent, it is possible that the band around 45 kDa may correspond to isoform 1 of CCNYL1: http://www.uniprot.org/uniprot/Q8N7R7
If you are unsatisfied with your results I would be more than happy to offer you a credit or a refund, but unfortunately we do not have a different antibody to this target at this time. Please let me know your original order number and how you would like to proceed and I will be happy to assist you further.
Answered on Mar 28 2012