Use 0.5 M Sucrose in your buffer, replacing 10 or 20 % glycerol. It works well.
Manish On Tue, Feb 23, 2010 at 3:07 AM, SERAH KIMANI <serah.kim...@uct.ac.za> wrote: > Dear all, > > Does anyone have an idea of something else that I can use instead glycerol > to maintain solubility of my protein? I have been having 10% glycerol in my > protein solution and this has helped me get crystals in like three different > conditions. However, I would want to get crystals of mutants-substrate > complexes, but unfortunately glycerol interferes with the binding of the > substrates to my protein. So, for the complexes to form, glycerol has to be > out (I have tested this using the wildtype enzyme both in the presence and > absence of glycerol). Now, in the absence of glycerol, I get a lot of > precipitation, and no crystals even after optimizing around the known > conditions with different protein concentrations. I have tried re-screening > for new conditions in the absence of glycerol, but I haven't found any > condition that yields crystals in the absence of glycerol. > > I was wondering if anyone might know of something else that I could use in > place of glycerol in my protein solution?? > > Regards, > > Serah > University of Cape Town > > ______________________________________________________________________________________________ > > UNIVERSITY OF CAPE TOWN > > This e-mail is subject to the UCT ICT policies and e-mail disclaimer > published on our website at > http://www.uct.ac.za/about/policies/emaildisclaimer/ or obtainable from +27 > 21 650 4500. This e-mail is intended only for the person(s) to whom it is > addressed. If the e-mail has reached you in error, please notify the author. > If you are not the intended recipient of the e-mail you may not use, > disclose, copy, redirect or print the content. If this e-mail is not related > to the business of UCT it is sent by the sender in the sender's individual > capacity. > > _____________________________________________________________________________________________________
