Protein purification. How to make function of recombinant protein? - (Feb/11/2011 )
I am senior student in bachelor degree. I have a problem. I purifind recombinant protein with Ni-chromatography in order to treat cell. However, i don' t know that the cell was died by my reconbinant protein or imidazole because imidazole may be affect the cell. Will I do it, if I does change into water or any compound which does not damage a cell? Could someone suggestme.
p.s. If you have paper, May you tell me too. Thank you for your kind.
did you also use urea when purifying the protein from the column?
if not then you can eliminate the imidazole by dialyzing the protein against a more compatible buffer.
if you used urea then you will have to refold the protein by stepwise dialysis to remove urea (and you can remove imidazole at the same time).
Thank for your suggesion. I use urea ,so i will try stepwise dialize on monday