Hello,

I am trying to purify a recombinant protein (~15kD) from Tobacco. I am using Agroinfiltration based expression system. Expression is high, but when it comes to purification, I could not get Protein of Interest rather more non-specific bands. I extract protein in extraction buffer (50mM Tris-Cl, 150mM NaCl, 20mM imidazol, 0.1% tritonX100, 1mM DTT, protease inhibitor), and after getting soluble protein, it is passed through the Ni-NTA column and afterwards washed in (PBS, 30mM imidazole, Protease inhibitor). Final elution is done in PBS+250mM imidazole.

Please suggest me some strategy based on your experience. If someone is expert in this technique, their cooperation will be highly appreciated.

Thanks in Advance