One of my research protein is bind to phospholipids, for elucidation of this binding mechanism I am trying to make lipid micelle for my interaction study through ITC. I am making stock of phospholipid (Phosphatidyl ethanoloamine) in chloroform and then vaporizing it through vaccum pump. After vaporizing, biofilm will be there on the wall of sample voil and then buffer is being added(Tris (20mM) pH 7.5, KCl (150mM) and Glycerol (10%). after adding this buffer, I did vortex and then sonication using water bath sonication. but after water bath sonication, I am not getting transparent solution of phospholipids. Is there any other suggestions,please let me know.
make liposomes. Phospethanolomie, any other phosphotidyl choline- egg or soy lecithin, choloesterol disolve in water and add to 60 degree water. liposome done in one hr.
With your method, your are producing liposomes. You cannot obtain limpid true solution in water with PE.
Hi Mr. Ajit,
I can suggest you to troubleshoot your process and see. Try to make a thin film as possible, this will help in forming small vesicles as possible.
Now second, reduce lipid concentration...again intention is to make thin film.
after you hydrate leave the solution overnight before you do sonication.
hope this may make some good difference.
Goodluck
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