So your protein is hopefully folded - that's good. Why would you want to
purify unfolded if you can purify native ? You can start with trying to
follow Quiagen's Ni-NTA instructions, for simple cases they work most of the
time. Let us know if you encounter something unusual along the way. Don't
forget to add 20-30 mM imidazole to your loading/washing buffer.
A.G.E.
<mrsankar_in at yahoo.co.uk> wrote in message
news:20030113104615.25934.qmail at ww02.hostica.com...
>> Hi all,
>> I have a His Tag protein overexpressed in E.coli and it does not go into
inclusion bodies. I want to crystallize the protein for which i need it in a
very high concentration and in very pure form. What way i can purify the
protein in Ni Column? Denaturing or native conditions?
>>http://biowww.net/mynews/tree.php?group_name=bionet_molbio_proteins&begin=0
