I would suggest the additive screen (Hampton Research) in general because it contains all of the afore mentioned compounds (by Puneet).
Also, have you thought about soaking the glycerol out of the crystal? It could work as long as you find a condition to stabilize the crystal without glycerol. I would suggest stepping down the glycerol concentration gradually while increasing the precipitant. Good luck, Kelly ******************************************************* Kelly Daughtry PhD Candidate Department of Physiology and Biophysics Boston University School of Medicine 590 Commonwealth Ave R 390 Boston MA, 02215 (P) 617-358-5548 ******************************************************* On Tue, Feb 23, 2010 at 8:20 AM, Annie Hassell <[email protected]>wrote: > > Serah-- > > We have used 1,2-propanediol to help with protein solubility. It has been > included it in our purification buffers and the final protein buffer in the > 3-5% range. > > HTH! > annie > > > > > Annie Hassell > Glaxo Smithkline > 5 Moore Drive > RTP, NC 27709 > 919/483-3228 > 919/483-0368 (FAX) > [email protected] > > > *"Puneet juneja" <[email protected]>* > Sent by: "CCP4 bulletin board" <[email protected]> > > 23-Feb-2010 06:35 > Please respond to "Puneet juneja" <[email protected]> > > > To > [email protected] > cc > Subject > Re: [ccp4bb] An alternative to glycerol in keeping protein soluble? > > > > > In general > > Osmotropes, chaotropes, amino acid, sugar , polyhydric alcohol, > detergents..... promote protein solubility > > Puneet Juneja > > On Tue, Feb 23, 2010 at 12:07 PM, SERAH KIMANI > <*[email protected]*<[email protected]>> > 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/*<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. > _____________________________________________________________________________________________________ > > >
