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Hi Iris, do you have any indications whether your oligomerization is concentration-dependent? If yes, you may be able to 'enrich' formation of your trimers if you do not exceed the critical concentrations leading to the larger oligomers. If you have access to DLS you could probably get some quick estimates of the concentration at which a particular oligomer is favored, or you can use analytical ultracentrifugation for a more accurate/detailed study. best wishes Savvas -----Original Message----- From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Iris Ben-Efraim Sent: Tuesday, October 03, 2006 10:15 PM To: [email protected] Subject: *** For details on how to be removed from this list visit the *** *** CCP4 home page http://www.ccp4.ac.uk *** Hi there, I have the following problem: I expressed a designed 67mer peptide/protein (7.8kd) which we believe forms coiled coils. On SDS-PAGE after crosslinking we see : monomer, dimer, trimer tetramer and pentamer. I'd like to get the trimeric form clean and so far I've used size exclusion chromatography superdex 75 column, it doesn't really work, the separation efficiency is practically non existent in this size range, also I suspect that the rod shape of the protein doesn't help here. Can anyone suggest a better separation method between the various multimeric forms? I'd like to get a homogeneous trimeric form. Thanks, Iris
