Darren-- Last year I attended the Membrane Protein Crystallization Course organized by Vivian Stojanoff at BNL. James Love talked about screening detergents using ASEC, and we have set this up in our lab now (not as elaborate a system used at the NYCOMPS). We use a "universal buffer" with a battery of 10-12 detergents and look for nice sharp peaks off the analytical size exclusion column. This has given us good starting points and clearly ruled out some detergents.
HTH! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) annie.m.hass...@gsk.com "Darren Hart" <h...@embl.fr> Sent by: "CCP4 bulletin board" <CCP4BB@JISCMAIL.AC.UK> 24-Mar-2009 11:22 Please respond to "Darren Hart" <h...@embl.fr> To CCP4BB@JISCMAIL.AC.UK cc Subject [ccp4bb] OT: Crystallisation compatible detergents Hello, We can make a nice 1:1 complex between two proteins that then gradually precipitates during storage at 4 degC. With 0.005% Tween-20 the complex is stable and does not preciptate noticeably. We have used this in buffers to measure the kinetics of binding by Biacore so it is clearly compatible with "functionality". Is Tween-20 at this concentration compatible with crystallisation? Is it worth giving a go, or a waste of time? Should we try and remove it, or reprepare the complex in an alternative detergent? If we need to try an alternative, what would people recommend? Thanks in advance for your suggestions, Darren -- ********************************************************************** Dr. Darren Hart, Team Leader High Throughput Protein Lab Grenoble Outstation European Molecular Biology Laboratory (EMBL) ********************************************************************** EMBL webpages: http://tinyurl.com/6xdltl http://tinyurl.com/667jrp Email: h...@embl.fr Tel: +33 4 76 20 77 68 Fax: +33 4 76 20 71 99 Skype: hartdarren Postal address: EMBL, 6 rue Jules Horowitz, BP181, 38042 Grenoble, Cedex 9, France **********************************************************************
