Question (31892) | Anti-Proteasome 20S LMP7 antibody (ab3329)

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Question

Dear Madam/Sir, We have recently purchased an antibody (20s LMP7) #ab3329 from Abcam but it does not seem to be functional. Me and my colleague have tried it with both milk and BSA blocking solution and it yields two non-specific bands much higher and lower than expected (20kDa) size. We also have purchase the Smurf1 and Smurf2 antibodies. The Smurf1 antibody works good but Smurf2 (#ab38543) antibody weakly gives the expected band but it shows also lots of sand shaped spots on the membrane and washing for long time does not solve this problem. I wonder to know if we could get replacement for these two new purchases from another batch of antibody. Thank you for considering our case.

Answer

Thank you for contacting us. I would like to ask you if you might send me the protocols used with these productsso that we can gather further information regarding the samples tested and the protocol used. Once we receive your response, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody. For our records could you please include the following details in your protocols for each antibody? What tissue type was used? How was the tissue prepared, including lysis buffer and conditions? Were the samples denatured? How much total protein was loaded per well? What were the running and transfer conditions and buffers? Did you use a loading control? Did you usethe recommended positive controls? What were the results of those controls? What blocking reagents did you use? You have mentioned that milk and BSA were used for ab3329, what were the concentrations of these in your block? What incubation times and temperatures did you use? What concentrations did you use the primary antibody concentration at? What incubation times, temperatures and reagents did you use? What was the secondary antibody? What were the concentrations, incubation times, temperatures and reagents used with this? Were different conditions tried, including different antibody concentrations and incubation times? What wash steps did you use? Which detection system did you use? I look forward to hearing form you soon.

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