What happens if you load your elution fraction into a size exclusion column? If
your protein of interest comes in the void volume together with most of its
contaminants, you'd better test a different construct, and that's much more
than only changing the tag from N- to C-terminus. Sumo and GST
Hi Lisa,
Have you compared your yield of purified protein of soluble protein per gram of
pellet, 4M Guanidine solubilized pellet (wash and elute from Ni column with 8M
urea .5M imidazole..otherwise the gel with run crappy), and and total protein
in the pellet on a page gel? The main reason I
Since there is a real possibility that it is I2 but most of the crystals are
pseudo-merohedrally twinned emulating I222, this is a case where I would
recommend keeping the I2 setting with beta close to 90 degrees rather than
converting to the more conventional C2 cell. Most modern programs (and so
It probably is I2 (== C2) so it would be better to re-integrate in C2 in
Mosflm, as the true beta angle may not be precisely 90deg
The CCs and R-factors are not very good, so it is worth looking at the scores
for the individual 2-fold axes output by Pointless, to check that one of the
axes real
