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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