Hi James, just out of curiosity. Did IZIT convincingly color the crystals? We have recently seen that crystals grown in high concentrations of MPD do not color with IZIT. This phenomenon was documented in Eckert et al. Acta Cryst. (2003). D59, 139-141.
Best regards Savvas ---- Savvas Savvides L-ProBE, Unit for Structural Biology Ghent University K.L. Ledeganckstraat 35 9000 Ghent, BELGIUM office: +32-(0)9-264.51.24 ; mobile: +32-(0)472-92.85.19 Email: savvas.savvi...@ugent.be http://www.lprobe.ugent.be/xray.html From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of james09 pruza Sent: Saturday, October 17, 2009 7:44 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Phase separation to crystals Dear CCP4bbers, Surprisingly, only At pH 3.6 the needles are appearing after 3 days but they are very thin and long rods. I have checked it running SDS-PAGE as well as using IZIT dye . Since the proteins has already 50mM of NaCl so I added 100mM of NaCl in the crystallization mix. The crystal appeared at 70% of MPD. Bunch of florets appears without NaCl at the same condition. Will the Additives like Ethanol or 2-propanol help in this condition? Should I not use NaCl in the mix? Changing temperature from 4degree to 25 degree will help? Please advice. Thanks a lot. James E-mail message checked by Spyware Doctor (6.1.0.447) Database version: 6.13500 http://www.pctools.com/en/spyware-doctor-antivirus/
