Would you consider purifying the proteins from a cytoplasmic extract instead of from the medium?

Hi Adam,

Thank you for your reply. I would consider, however, I have purification issue with extracting from cells.  

You could begin with metal affinity chromatography, thanks to the His tag, but this would probably not be sufficient to get the protein fully purified from Sf9 cells. You would then have to follow up with at least one more chromatography step. Affinity chromatography on a heparin-Sepharose column based on binding of fibronectin to heparan sulfate is one approach. Gel filtration chromatography is another, since the protein is quite large.

Dear

Your protein is intracellular or extracellular.

Thank

Thank you Adam

 Hi Marius,

I expect to secrete the protein into the media as I have included signal peptide at the n terminal to my chimeric protein of interest. However, lower mwt proteins were able to secrete but 75 KDa and above are not. Cell lysated tested on western shown interested proteins with expected size intracellularly. So my task now to purify from cells