Protein Purification from insoluble cytoplasmic fraction

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apees7k
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Protein Purification from insoluble cytoplasmic fraction

Post by apees7k » Mon Nov 13, 2006 5:08 pm

I am expressing a DNA interacting protein. The gene is cloned in pET28b vector and all of my protein is going to the insoluble cytoplasmic fraction (cell debris or inclusion bodies). I checked it by performing western that my protein is overexpressing but I know from my experience that it is very hard to get a functional protein out of it. As I am trying this type of protein (DNA interacting) for the first time so can someone suggest how can I proceed and what can be the best strategy to purify and achieve a functional protein. I would appreciate if someone can suggest a kit or whatever relating to the purification. (Just to let you know that the recombinant protein contains a 6XHis tag).

Thanks
AP

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