Hi all,
I’m trying to purify a filament protein (extracellular) and I have encountered a huge membrane contamination most likely due to the bleb from my bacteria. I have attempted sucrose gradient, but my protein kind of run at the similar gradient with these lipids. Also, I tried detergent, but it could break my protein apart due to the hydrophobic core they have. I also tried gel filtration, the membrane was all over the place and co-eluted with my filaments. I am wondering what would be a better purification strategy to remove membrane contamination? If no, is there any non-fluorescent dye for membrane which can indicate the membrane if I want to optimize sucrose density?
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