An added benefit of EDTA is that it inhibits some proteases - for one
of our wimpier proteins, spiking each fraction collector tube with a
little EDTA before running the Ni column really helped reduce keep
the sample in one piece.
Phoebe
At 01:18 PM 2/19/2008, Sophia Tsai wrote:
Hi,
Agreed on this. I used to have issues with aggregation due to the
Nickel being stripped off the column (happens with elution in
imidazole). Adding 10mM EDTA to the elution immediately AFTER it has
come out of the column will chelate the Ni++ and prevents
aggregation (at least in my case). Afterwards, I use gel filtration
to remove the Ni++, as well.
Hope that helps!
Sophia
On Mon, Feb 18, 2008 at 4:48 AM, Ngo Duc Tri
<<mailto:[EMAIL PROTECTED]>[EMAIL PROTECTED]> wrote:
Hi,
I used another way to deal with this problem. You can try to elute
your protein with the buffer containing 50mM EDTA (You need at least
10CV to elute completely). Then use gel filtration to remove the Ni.
I applied this method with two proteins and it showed good results.
Good luck!
TriNgo
---------------------------------------------------------------------------------------------------------------------------
Phoebe A. Rice
Assoc. Prof., Dept. of Biochemistry & Molecular Biology
The University of Chicago
phone 773 834 1723
fax 773 702 0439
http://bmb.bsd.uchicago.edu/Faculty_and_Research/01_Faculty/01_Faculty_Alphabetically.php?faculty_id=123
RNA is really nifty
DNA is over fifty
We have put them
both in one book
Please do take a
really good look
http://www.rsc.org/shop/books/2008/9780854042722.asp
