antibodies from inclusion bodies
no.email at thanks.to.spam.net
Wed Apr 23 16:39:59 EST 2003
In article <20030423185347.12208.qmail at ww02.hostica.com>, humphreyt at agr.gc.ca wrote:
>My recombinant protein is strongly expressed in inclusion bodies in E coli. I
> have heard that purified inclusion bodies can be directly injected into
> rabbits for antibody production without the need for solubilising and
> refolding. Has anyone tried this and / or have any hints and tips for me?
Yeah, it's been tried many times. It's quick and dirty. Invariably,
such sera work but show a lot of cross-reactivity/high background.
Considering how easy it is to purify 6His-tagged protein under
denaturing conditions (or even non-tagged by ion exchange in the
presence of urea) and use it in immunizations, I personally
find there is no excuse for using crude inclusion preps in
immunizations. No refolding necessary - you can inject unfolded
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