Hi,
I'm purifying my protein with the His-TAG-method.
My protein, the ATCase of pyrococcus abyssi consists of two different
subunits. The His-Tag is attached to one of them.
After elution with Imidazol (100 mM Imidazol), I concentrated my protein
and put it on gel.
On SDS gel it looks very nice and pure, on a native gel on the other
hand, there is a smear in which I can distinguish one band.
The native gel is a 6 % Tris-glycine gel.
Is it possible that the high imidazol concentration has an influence on
the migration in my native gel.
Is it possible that because of the His-Tag the protein forms higher
molecular weight aggregates, dimers?, Trimers?
Can I remove the Imidazol just by dialyzing?
Thanks in advance
Sigrid
Sigrid Van Boxstael
VUB MICROBIOLOGIE
E. Gryzonlaan 1
1070 Brussels
Belgiuml
