[Protein-analysis] Glycerol during protein purification and protein oligomerization state

Ligong Wang via proteins%40net.bio.net (by ligongw2004 from yahoo.com)
Tue Jul 24 09:39:33 EST 2007


I have not used, during protein prep, more than 10% glycerol.
 However, I did follow one protocol, at the end of the prep, to store the
 purified enzyme in 50% glycerol at -80 degree frozen. I guess the percentage
 of glycerol is not much a concern as long as your procedure/instrument
 can endure the high viscosity/back-pressure. I am wondering though
 about your rationale for increasing the glycerol. Are you suspecting
 hydrophobic, non-specific stickiness of the protein be the culprit for your
 problem? Is there other reasons that might cause the issue you observe?
 disulfide bond formation? ionic interaction? protein unfolding? etc.
 Practically, there are many steps/details that can be altered from one
 prep to another, even though one may not realize it. I had many such
 experiences, personally and from colleagues. It sure helps to back track
 every little details of your operation to trouble-shoot. Hope you can
 get over this hurdle soon. Good luck.

Ligong Wang
Yale University

       
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