"Debashis Mukhopadhyay" <debmukh at scripps.edu> wrote in message
news:Pine.SGI.4.20.0301141518540.199543-100000 at signal.scripps.edu...
> Hi,
>> I am using Ni affinity column to isolate my protein which came out
> successfully with Imidazole. However, I found a lot (~100 mg) of protein
> still sticking to the column and I eluted them out by stripping off the
> Ni+2 from the column using EDTA. So, this solution now contains my protein
> + (may be some contaminant) + Ni and EDTA and is colored blue (as
> expected). At this point I wonder if there is any way / standard practice
> to separate the protein from Ni+2, so that I can re-utilize it.
>> Any help would be appreciated.
>> Regards,
> Deb
It's not very likely that the protein would be OK. In my experience, strange
things happen to proteins exposed to soluble nickel salts, and Ni tends to
persist even after thorough cleanup. You can try, however - simple desalting
(size-exclusion) should do the job. After desalting, remaining nickel can be
sometimes scavenged using chelating resins.
A.G.E.
