Hi! Maybe you protein is present as soluble microaggregates and gets stuck on the resin, without really 'binding' to it. I would spin the lysate in an ultracentrifuge 1h @ 50 k or so, to see if there is anything left in the supernant afterwards. And then make decisions based on that outcome.
best, sebastian > On 15. Sep 2017, at 13:53, Narayanan Ramasubbu <ramas...@sdm.rutgers.edu> > wrote: > > Hi. We are working on a periplasmic protein that breaks naked glycans in > peptidoglycans. There is truncated structure available but our target is the > full length protein. The difficulty us that it strongly binds to the resin > with or without his.tag. Changing the resin to acrylamide did not help. > Has anyone come across similar problem and how was it resolved. > The pdb structure is the catalytic domain and mussing a region that, in my > opinion, binds to the resin. > Thank you in advance > Sent from my iPhone
