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These antibodies not working correctly in Western blot.
Asked on Mar 28 2012
Thank you for your calls earlier this month and for your patience.
Have the replacement vials of ab53060 and ab2451 provided any new results?
I've reviewed the protocols used to test these antibodies, and I didn't notice anything particularly unusual. The only difference was thatBSA was used as the blocking agent for the E2F antibodytestings, while the p107 antibody was tested on a membraned blocked with Blotto (milk based). Some antibodieswork better with a certain blocking agent,though I can't be sure that thisis a cause of the absent bands in this case.
The lab that tested ab2451 did report seeing a band between 55 and 60 kDa in their samples, however this was in addition to a strong specific band at the expected molecular weight. This extra band is believed to be a cleavage fragment, but no further characterization has been done on the band.
Please let me know if there has been any improvement with the results using the replacement antibodies, and if you would like to try a replacement for the E2F3 antibody I will be happy to send one.
I look forward to hearing from you. Let me know if you have any questions or if there is anything else that we can do for you.
Answered on Mar 28 2012