Covalently cross-link a GST fusion protein to GSH agarose?
IAmNotFredSoPleaseDontSendMeAnyEmail at hotmail.com
Tue Mar 6 01:38:31 EST 2001
Thanks Rick. But Pierce has a LOT of cross-linkers. To which one were
you referring? Or does the special kit have a name??
I don't want to do the prep cell thing. We found some polymers of our
GST fusion protein at various MWs thru the whole gel - ID'ed by
MALDI-TOF. Its these sort of artefacts I want to particularly avoid.
Rick Thorne wrote:
> Dear Fred/Phil/whatever \yournameis,
> Pierce sell just a kit to do as you describe. I have not tried this kit but
> I have already tried the same X-linker myself (before I saw the kit ) but I
> was not all that impressed by the results. Maybe they have optimised the
> protocol. You should also consider that the X-linker might screw up your
> interaction(s). I have also tried other means to get rid of the fusion
> protein but there is so much of it relative to the binding proteins it
> causes major headaches. My solution is to run the whole damn eluate on a
> BioRad 491 electrophoresis prep cell which can seperate things by size (all
> of our bp are bigger than the fusion).
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