I plan to perform cell fractionation and subsequent protein localization in E. coli but I want to disrupt my cells gently so that the protein complexes remain as stable as possible. Sonication is fast and allows small scale lab work, but I suspect it may disrupt weak protein-protein and protein-lipid interactions and alter results on protein localization. Could French press, although more tedious, provide a more gentle disruption? I have read nothing against sonication for protein localization studies, but I feel in most papers French press is preferably used for this purpose. Could anyone illustrate my doubts?
Thanks for your reply.
As I plan to obtain membrane samples I think chemical, disruptor and freeze/thaw-mediated lysis may not be efficient enough to obtain significant uncontaminated membrane yields with my fractionation protocol. I have already tried a cell disruptor with no satisfactory yields for membrane samples. I could try to optimize it though ...
I will consider your suggestions, thanks again!
I again appreciate your reply.
Actually, my fractionation protocol is similar to yours, with some additional steps including sodium carbonate treatment. I will check your reference, I may take some ideas from yours.
Thanks again.
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