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
