I prepared cell culture with phospolipid of interest.
However, in extracting it I shouldnt use any kind of solvent.
What techniques would you use that dont have liquid phase for extraction of those lipids?
Hi Stefan, I'm not sure what experiment you want to do, but if you just want cell membrane lipids without proteins, I would just use proteinase K and/or trypsin to digest all of the proteins associated with the membrane. Also do occasional sonication to expose inner membrane proteins to the proteinases.
Membrane lipids have a high CMC, so you can dialyze away cleaved peptides using a 3-8K dialysis bag. Proteinase K and trypsin are easy to inactivate.
@Paula what is "supercritical fluid extraction with carbon dioxide"?
Super critical CO2 is https://en.m.wikipedia.org/wiki/Supercritical_fluid
Steingrimur I'm trying to crystalize biomoleciles outside water or liquid medium. Using powdered trypsin and other enzyme can work?
Are you suggesting me to purify first lipids and then crystalize?
Hi Stefan I thought you just wanted a natural phospholipid preparation without all the proteins.
I don't know what biomolecules you are trying to crystalize or how to help you on that.
natural phospholipid preparation without solvents. and water... extraction that not use liquid phase
I'm still not sure of what you are trying to do so I don't know if this helps, but here is a way to use triton X-114 phase separation to isolate membrane lipids and membrane protein.
http://www.jbc.org/content/256/4/1604.abstract
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