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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