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 <[log in to unmask]> 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
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