protein precipitation

James Fethiere james at mpimf-heidelberg.mpg.de
Thu Aug 20 12:27:35 EST 1998


Hello, 

I was just preparing to purify a soluble kinase from the cytoplasm of
sf9 cells but ran into a little problem.  After polytron, and high speed
centrifugation, the published method indicates that the supernatant
(soluble fraction) should be completed with a HEPES buffer pH 7.2
containing EDTA, and 0.02% Triton X-100 before going to an S-sepharose
column. Well, the prepared buffer looks very clear, but as soon as I
dilute the protein fraction with this buffer, everything becomes cloudy,
and a light precipitate forms.  I don't see any reason for this.  Any
ideas????

Thank you

James



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