Did you filter your lysate through .45 then .22 filters?
cheers
b
Quoting James Stroud <[EMAIL PROTECTED]>:
Try refolding before purification.
On Mar 2, 2008, at 11:47 PM, Tim Gruene wrote:
Hi all
sorry, for offtopic query...
I am trying to purify my protein by Ni-NTA affinity chromatography. After
sonication as i centrifuge bacterial lysate, soon after 10 min
whole lysates
get precipitated during loading on the column and some time it remain
soluble too. if i get purified through the column without
precipitation, it
gets precipitated during dialysis.
I have tried lot, by chnaging buffers, increasing salt or deacreasing salt
or no salt at are helpless.
I do purifiaction in cold room.
can any one suggest some solution?
Thanks in advance.
NSH
--
James Stroud
UCLA-DOE Institute for Genomics and Proteomics
Box 951570
Los Angeles, CA 90095
http://www.jamesstroud.com
