I am afraid that you might have your protein in an unfolded state and
thus, trapped by the chaperon.... At least this is what I have
experienced in the past... If the protein is properly folded but it
gets trapped by the chaperon due to hydrophobic interaction, washes
with ATP, Mg2+ and K+ generally works...
I would start thinking in another expression system...
Good luck,
Maria
Quoting Gloria Borgstahl <[email protected]>:
We have had good luck with making a protein soluble using the Artic Express
bacterial cell line BUT we can't get rid of the chaperone that copurifies.
We have tried adding ATP, MgCl2 and potassium to lysate and extensive
washes and this releases a bit of the chaperone. Has anyone solved this
problem? We have heard of adding denatured proteins to the lysate to
confuse el chaperono. Does this work?
Many thanks in advance for a advice, Gloria
