Imidazole + enzyme activity - (Jun/28/2006 )

Hey all,

Does anyone know whether or not imidazole has any affect on enzyme activity? My protein is currently contained in a buffer containing imidazole and am unsure whether or not I need to dialyse it out. The buffer contains 0.02M sodium phosphate, 0.5 M NaCl and 0.5M imidazole. If I need to dialyse, what should I dialyse against?

Thanks for any help you could provide.

-btjd-

We use 250mM imidazole and it doesnot have any effect on enzyme activity. Not sure abt 0.5M.

-scolix-

am not sure about imidazole affecting activity, but 500 mM of NaCl might, I read something about desalting your protein in some amersham columns (P10 if I can remember).
I was also thinking of eluting the protein in a buffer more proper for its activity or further utilization

-tertu-

We use 500mM Nacl along with 250mM imidazole and we didnt have any problem with enzyme activity.

-scolix-

I dialysed against PBS, I don't know where it can be desalt by column of millipore?

-knewman-

Check this out
the columns are PD10 from amersham
http://www.mdyn.com/aptrix/upp01077.nsf/Co...273&zone=Labsep

QUOTE (knewman @ Jul 1 2006, 01:49 PM)

I dialysed against PBS, I don't know where it can be desalt by column of millipore?

-tertu-

我使用了millipore的柱子, 脱盐效果很好, 而且速度很快, 节省时间

-knewman-