Question (32054) | Anti-Proteasome 20S LMP7 antibody (ab3329)

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Question

Dear Madam/Sir, Thank you very much for cooperation in resolving our problem with the antibodies! I am attaching a film image of the lysates from mouse endothelial cells (first 4 lanes) and Pericytes (lanes 5-8) blotted with Anti-20S core Ab from abcam. The higher band is about 52 kDa and the lower band is almost 5 kDa. I could not see any other band with size 20 kDa as expected. The smurf2 antibody is now working by increasing the concentration to 1/100 and washing 5X with TBST. I am attaching the ordering information as you requested. Please let us know when we could get the new batch of the anti- 20S Ab. Thank you, Best regards,

Answer

Thank you for sending me this image. This data will be used to keep records of how our products are performing and for internal invetigation if needed. I noted that all tissue lysates that you have used have given the same non-specific signal. However I was wondering if you had tried a positive control lysate with your experiments. I ask this because according to SwissProt, in mouse tissue,this protreasome is expressed in spleen, thymus, lung, liver, heart and, at a very low level, in kidney.However it is not expressed in brain nor testis. My concern is that whilethe antibody may be recognizing non-specifically it may still be performing as there is not expression in the brain. This information is in the general annotations on this page: http://www.uniprot.org/uniprot/P28063 I would highly recommend using a control tissue lysate such as ab7937 Spleen (Mouse) Whole Cell Lysate - normal tissue ( https://www.abcam.com/Spleen-Mouse-Whole-Cell-Lysate-normal-tissue-ab7937.html). If you have any questions please do not hesitate to contact me.

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