Posted 21 July 2011 - 07:04 AM
I ordered the production of FGF-B in E. coli and I obtained the purified protein in PBS plus 8M urea. I was told the protein went to inclusion bodies and thus, they had to used urea to dissolve the protein. The point is that I need the native protein. Does anyone know of a good protocol to refold this protein or a related one? If not that specific, any more general protocol?
I was thinking of dialysis... is it possible to progressively reduce the urea concentration? Should I use glycerol in the dialysis buffer as well?
Thanks a lot in advance!
Posted 21 July 2011 - 04:41 PM
Posted 21 July 2011 - 10:21 PM
Returning to refolding, if it has S-S intramolecular you have to add reducer to your urea solution, and having present that intermolecular S-S (reaggregation) could be formed if the concentration is too high or the pH is near the isoelectric point . Buena suerte