peter.cherepanov at uz.kuleuven.ac.be
Fri Dec 14 04:59:21 EST 2001
some proteins renature better when you do a step-dilution rather then slow
dialysis, it is also much easier.
Try adding it drop by drop into a buffer minus denaturant. You might see it
precipitating, only part might be soluble. If it works well, it will start
precipitating only when protein concentration rises above some limit. Mix
after each drop. Do it on cold (it reduces hydrphobic interactions). For
some proteins (that have lots of charged residues) adding NaCl (0.5M) might
help, for very hydrophobic proteins that will likely only worsen
and Cys residues- do you include DTT ?
and of course, some proteins will not be soluble even in their native forms
without a detergent ...
Does CBD work in denaturing conditions ?
does it also have His tag ? (I heard that some vectrors with CBD have it)
<- then may be renaturation on the column might work better.
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