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Try buffer and additive screening. If some lab nearby has a real time
PCR machine
you can screen 96 conditions in an hour with the Thermofluor method.
Check out:
Ericsson UB, Hallberg BM, Detitta GT, Dekker N, Nordlund P.
Thermofluor-based high-throughput stability optimization of proteins
for structural studies. Analytical Biochemistry 2006 Aug 10; [Epub
ahead of print]
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?
db=pubmed&cmd=Retrieve&dopt=AbstractPlus&list_uids=16962548&query_hl=1&i
tool=pubmed_docsum
Good luck,
Martin
On Sep 22, 2006, at 9:50 PM, Jenny wrote:
Hi All,
I'm trying to purify a protein which is expressed in soluble
form.It's a His-tag protein, so when I run His-column, it's really
easy to clog.In the next step size exclusion purification, the
protein come off at the void volume, which suggests that the
protein is severely aggregated.Does any one have any suggestions
that I can modify the purification condition so as to avoid
aggregation problem?Or can I add GuHCl during His column
purification so that it won't clog so easily?Thanks a lot.
Jenny
