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Thank you for the mail. First of all I have to confirm once again that your antibody produced many unspecific band, but no specific one. If as you said, the blocking was too long, we should not have got any bands, but our problem is too many bands although we blocked overnight, which works very well for other antibodies. Do you think only your antibody targeted protein was lost? Your suggested incubation time at 4°C overnight, I don´t think it will become better (according to my 20 years´s experience). I have read data sheet from abcam once again, you have no any record of the antibody to detect the full length protein, you have just used it to recombinant p75, could you please provide any kind of record of related tests? All the best!
Asked on Sep 05 2012
Thank you for your reply.
I appreciate the time you have spent on these experiments in the laboratory and it is disappointing the results have not been successful so far. As discussed in my previous email, although other antibodies in your laboratory have worked well under these conditions, all individual antibodies will require individual optimisation. It is not unusual to have to do a few optimisation experiments with a new antibody. We often find that suggesting some scientifically thought out optimisation tips improves the results. This is obviously a preferable outcome for the customer.
However, having revisited the datasheet, it looks like this antibody has not been tested on the endogenous mouse protein so far. I fear that this may be the cause of the problem.I am therefore happy toissue you the replacement with the ab96428 Anti-Ndufs1 antibody.
Usually, our replacements are also covered by our Abpromise, but ab96428is only to predicted to work with mouse samples, and that would mean in case of a failure the experiments are not covered. Checking our website, we have another antibody which is tested to work in mouse: ab104246. Considering the trouble you had with the previous product, I will ask the lab if they can provide a WB image for the mouse lysate to be on the safe side.
Please let me know if I should still send you the replacement of ab96428now or if you would prefer to wait for the answer of the ab104246case.
Answered on Sep 05 2012