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) annie.m.hass...@gsk.com "Puneet juneja" <junejabiot...@gmail.com> Sent by: "CCP4 bulletin board" <CCP4BB@JISCMAIL.AC.UK> 23-Feb-2010 06:35 Please respond to "Puneet juneja" <junejabiot...@gmail.com> To CCP4BB@JISCMAIL.AC.UK 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 <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. _____________________________________________________________________________________________________
