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Thanks for your response. For some reason the email I sent a couple days ago bounced back. Please see attached an overlay of the ECL image with the Marker image. It definitely seems like there are two bands (between 75 kD and 50 kD) that correspond with the BACE1 protein. Specifically, one at approx 70 kD and one at approx 56 kD. Based on the data sheets you sent me, the 70 kD band is probably the glycosylated form and the smaller approx 56 band might be the unglycosylated form. It is strange that the mouse lysate appears in the 100 kD area, but there seems to be a faint band in the 3T3 lanes at approx 56 kD. I need to re-run this in the middle of a gel to be sure, but I need more lysate.
I'm not ready to jump ship and go with a different company. Is there another positive control you'd recommend? Also, since the peptide hasn't worked, I'm guessing the batch was somehow bad. I also think I should try a new BACE1 antibody along with the ab2077 I still have. So, can you please send me the following: 1) peptide ab7883, 2) either a different positive control or more of the mouse lysate 3T3 (at least 40ul), 3) ab108394 (since perhaps a monoclonal Ab would decrease the non-specific bands I'm seeing).
Asked on Jun 15 2012
Thank you for your reply.
I am sorry about the issues you have been having with ab2077 in western blot. I cannot send you all of the reagents that you are requesting, I can either send 1 free antibody as a replacement for ab2077, or I can process a refund for the cost of the original purchase.
Please let me know how you would like to proceed.
Answered on Jun 18 2012