Reza,
someone might have mentioned this, it takes longer time to cool down larger
culture, we turn down the temp of the shaker when OD is about 0.4. For some
protocol, we even use ice to cool the flask down before induction. You
might also want to consider a lower induction temp, like 16degC.
Hi Jerry,
We have used 0.05% Triton X-100 in our wash buffer to wash the protein
on column (Ni-IMAC for His-tagged proteins and MabSelect for
antibodies). 20 column-volumes wash followed by 20 column-volume wash
without detergent is typically enough to reduce the endotoxin
significantly.
I would
