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I recently tried using Abcam antibody ab5295 (anti-RanBP9) for western blotting, with limited success. I've attached a summary of the procedures and results as a powerpoint file. Perhaps with some further optimization, signal to noise could be improved, but I figured I'd send what I have now anyway. Please let me know what you think.
Asked on Jun 25 2004
Thank you for getting back to us and for sharing your results with us. As you probably know, this is a Fast-track antibody and it has not been fully characterized. We have read through your protocol and would like to make the following suggestions/comments: 1. Try to use PVDF membrane. Generally, we find enhancement of transfer of proteins on PVDF membranes compared to nitrocellulose membranes. 2. We would also suggest diluting the antibody at 1:500 and incubating the filter overnight at 4 oC to see if the signal is getting stronger. 3. If you have enough material, try to load at least 20-30 ug protein/lane. Please do let us know how you are getting on with this antibody. We encourage customers to feed their results back to us, whether positive or negative, and make all information about each of our products available to researchers. In order to be able to send you a USD15/EUR15/GBP10 Amazon gift voucher, we would like to ask you to re-submit your review through the on-line datasheet which is the normal way for submitting feedback information. Even if you have already sent us your results to this e-mail address firstname.lastname@example.org, we cannot resend your message as a review on-line - we have no right to do so. We look forward to hearing from you soon.
Answered on Jun 28 2004