Hi Rhys, I hope you are well. The JOVE article for bicelles is nice, plz check the video. There is a table in the text section showing some structures that originated from crystals grown at 4-7 % bicelle conc. using not only the DMPC/CHAPSO mixture... The other parameter that everyone tries is of course temperature which has effects on the crystallization process, viscosity & phase diagram (because of detergents, and more complicated due to the additional components in this case), etc. Probably you know..: bicelles' term = bilayer + micelles... there's a thousand parameter one can adjust, and screening is the best way around it for now when "used as a method". Once I read that a team obtained great crystals with it after six months at 4 °C... So many advices/logic in our field are "time dependent" :) Once you have the bicelle stock which can be stored for long time.. setting up the trials then is as easy as normal/classic methods. Not much experience, but for similar lipid-rich methods for example, minor amounts of detergent can strongly change the phase behavior... We might have heard of too many hypotheses, but some say that anything (more than one phase) that has a minimum and particular arrangement// i.e., is a bilayer etc, may result in crystals... Best wishes toufic el arnaout
________________________________________ From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of RHYS GRINTER [r.grinte...@research.gla.ac.uk] Sent: Friday, December 13, 2013 3:54 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Bicelle Crystallisation Hi All, I'm thinking of embarking on some crystallisation of a membrane protein in bicelles in the new year. In the methods I've read you simply take your protein in detergent and add the bicelle mxiture (Chapso+DMPC) to your protein allow the solution to equilibrate and set up your screens. What I was wondering is will the original concentration of detergent in your protein sample, effect the likelihood of crystallization (i.e. by changing the structure of the bicelle)? If so do people generally seek to minimize the detergent concentration of the sample before setting up the bicelles. Cheers, Rhys
