lysate reduced Ni resin

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lysate reduced Ni resin

I ran lysate (from e.coli)  through a HisTrap ff column and the resin
turned a purple/brown color and my yield was rather lower than usual.
Neither the wash (lysis buffer) nor elution buffers had any effect on the
resin when I equilibrated the column.  Also, the pH of the lysate matched
the lysis/wash buffers.  Has anyone experienced this?  This is the first
time this happened during the purification of this particular protein
(this will be the 4th prep)...

Any help would be appreciated!!!


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