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
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