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Dear Julia,
Does your protein contain a coiled-coil? If yes, it does not mean that it is aggregated - it simply cannot enter the pores in the GF media.
I would be cautious here. We are working with a number of coiled-coil fragments which are most typically about 80 residues long (some are longer). A coiled-coil of 80 residues is a 12nm long rod. They are expressed with an N-terminal 6xHis tag which is cleaved off after the purification of the Ni column. As the final step, we use gel-filtration on Superdex 75. Most fragments apparently do enter the column and yield a decent peak (these are double-stranded coiled coils). However, we did see aggregation with some of our fragments. In this case there is indeed a peak or a family of peaks close to the void volume. Best wishes, Sergei -- Prof. Sergei V. Strelkov Laboratory for Biocrystallography Department of Pharmaceutical Sciences, O&N 2 Catholic University of Leuven Herestraat 49 bus 822, B-3000 Leuven, Belgium Phone(work): +32 16 33 08 45 Fax: +32 16 32 34 69 Phone(home): +32 16 23 19 06 Mobile: +32 486 29 41 32 E-mail: [EMAIL PROTECTED] Disclaimer: http://www.kuleuven.be/cwis/email_disclaimer.htm
