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[ffaucher]

Hi,

I'm working on a 40Kda His-tagged protein and I have too much contaminant protein after elution on Ni-Nta column.

I tried to wash with increased concentration of imidazole until my protein is eluted, but contaminant protein are eluted too.  I tried gel filtration but contaminant protein still here.  I think that there is too much protein interaction betwen my protein and contaminant.

How to reduce protein interaction?

I tried chaotropic salt (NaSCN) and increased salt concentration. with encouraging results but it's not enough.

Thanks

Fred F