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Iris Ben-Efraim wrote:
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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
What about ion exchange ? You should be able to get all your species
separated. Try runs with small amount of protein first and vary pH,
buffer and the steepness of the gradient.
Juergen
--
Jürgen Bosch
Howard Hughes Medical Institute and
University of Washington
Dept. of Biochemistry, K-426
1705 NE Pacific Street
Seattle, WA 98195
Box 357742
Phone: +1-206-616-4510
FAX: +1-206-685-7002
