Generally it might not be the best thing to do, especially if you have
traces of transition metals in your solution. You can expect some
hydrolysis if you decide to autoclave concentrated imidazole solutions -
whether this would affect the behavior of your IMAC - I can't predict
w/o trying. The one time I autoclaved 300 mM imidazole at pH 8 it turned
slightly yellow.
My guess would be that even if some imidazole decomposes/polymerizes,
there will still be enough of the intact compound left to work with. I
am not sure what these decomposition products might do to your protein,
though.
If you were to autoclave things, I'd say that pH 5 would be safer than
pH 8 but that's just a guess. Since most IMAC procedures use pH 8, you'd
still have to adjust your pH in the end so autoclaving at pH 5 makes no
sense.
What's wrong with sterile filtration, anyway ?
Artem
Michael Witty wrote:
>> Dear All,
> what are our opinions on autoclaving imidazole in buffers for
> Ni-NTA? I think we shouldn't because this would cause imidazole
> decomoposition. Is there any experience out there? Mike.
--
|Dr. Artem Evdokimov Protein Engineering |
| NCI-Frederick Tel. (301)846-5401 |
| FAX (301)846-7148 |
|eudokima at mail.ncifcrf.gov |
|http://www.ncifcrf.gov/plague |
