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I have purchased the anti-PABP antibody (ab6125) and used it successfully in western blot experiments. Recently I have attempted to use it in antibody inhibition experiments with RNA and have found that the RNA is being degraded during the experiment. I was under the impression that these antibodies were highly purified and therefore should be free of ribonucleases. It would be greatly appreciated if you have knowledge about the purity of the antibodies generated (if it is lot dependent) and if abcam or any other company guarantees their antibodies to be nuclease free.
Asked on Jul 30 2007
Thank you for your enquiry. I have received information from our laboratory. The purification steps are not done aseptically (it's impossible) and so whilst I guess there could be some bacterially derived endonuclease, I think it unlikely as all our columns are stored in ethanol and all the tubing etc washed extensively before each run. Moreover, even if there were any endonucleases present, I would be surprised if they bound non specifically to protein G and so I wouldn't expect to see them in the end product. This is substantiated by the fact that we don't see any low (12-14Kd) MW bands in the preps when we run SDS-PAGE gels. Whilst we filter sterilize the preps prior to despatch, I guess if the antibody has subsequently been handled non-aseptically at any point, this could introduce bacteria which could produce endonuclease, however I think it unlikely. I hope this information will be useful to you. Should you require any further information, please do not hesitate to contact me.
Answered on Jul 30 2007