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Why were the cells harvested immediately after IFNalpha treatment? Do you have a reference explaining that this is necessary to maintain the integrity of the lysate proteins? If I am using the lysate in Western blot, do I need to handle them differently than untreated lysates?
Asked on May 10 2011
Thank you for your phone call the other day. I have heard back from the production lab regarding the IFN alpha treatment of the cell prior to lysis, and I received the following response: "The treatment with IFN alpha has no effect on the stability of the protein or of the extract. When we say 'cells were cultured in medium supplemented with 500U/mL interferon alpha for one hour at 37°C immediately prior to harvesting' it only means that there was no gap between the treatment of the live cells and the preparation of the nuclear extracts. The cells where not frozen or anything like that." I hope this information is helpful, but please let me know if you have any further questions.
Answered on May 10 2011