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On several occasions we've had to use 5-10% glycerol in the protein. Remember to add the same to all your screens since otherwise the vapor diffusion often works 'backwards' (i.e. the drops get fatter with time). Artem -----Original Message----- From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of Rob Gruninger Sent: Friday, August 24, 2007 7:09 PM To: [log in to unmask] Subject: [ccp4bb] Off Topic:crystallization in the presence of glycerol Dear All, I am working with a protein that requires 10% glycerol throughout the purification to keep it soluble. I have been very worried that having glycerol in my protein solution when I am trying to crystallize it will prevent me from obtaining crystals. I am curious to see if others have successfully obtained crystals of proteins that required glycerol to keep them soluble and if so what concentration of glycerol could you use? Should I be concerned about this or am I worrying too much? Thanks for your input Rob -- Robert J.Gruninger B.Sc. PhD Student Department of Chemistry & Biochemistry University of Lethbridge, 4401 University Dr. Lethbridge, AB, Canada, T1K 3M4 Phone:(403)329-2746 Fax:(403)329-2057