aggregation in dialysis

Priscilla Too priscillatoo at cuhk.edu.hk
Wed May 29 07:12:02 EST 2002


How about dilute the protein before dialysis?



"Mariella" <parisi at fis.unipr.it>
???????:fad5fa87.0205090019.5692049b at posting.google.com...
> The buffer was 20 mM, but I tried also with NaCl 250 mM and glycerol 300
mM.
> I never tried detergents.
>
> > What's the salt concentration ? Did you try detergents yet ?
> >
> > A.G.E.
> > "Mariella" <parisi at fis.unipr.it> wrote in message
> > news:fad5fa87.0205072344.5b20a4eb at posting.google.com...
> > > I used continous flow dialysis against buffer (TRIS or Phosphate
> > > buffer PH 7) for changing buffer. The time isn't important.
> > >
> > > "Artem Evdokimov" <AEVDOKIMOZ at cinci.rr.com> wrote in message
> >  news:<9g_B8.58555$Ez5.14761633 at typhoon.neo.rr.com>...
> > > > Detergents ? Continuous flow dialysis ? Dialysis via
ultracentrifugation
> >  ?
> > > >
> > > > it really depends on the details - how long do you dialyze, against
what
> > > > (and from what) etc.
> > > >
> > > > A.G.E.
> > > > "Mariella" <parisi at fis.unipr.it> wrote in message
> > > > news:fad5fa87.0205070655.3e5eafde at posting.google.com...
> > > > > Hi everybody!
> > > > > I've a problem. The protein I study gets aggregated when I put it
in
> > > > > dialysis. I've tried many things: to change the buffer, collodion
> > > > > bags,
> > > > >  to add EDTA or Glycerol or Salt. But nothing.
> > > > > By X-ray it seems that nothing binds it, and so I've rejected the
idea
> > > > > that
> > > > > the protein loses a ligand in dialysis.
> > > > > The protein is the porcine Odorant Binding Protein (a Lipocalin):
its
> > > > > analogous one ( that one extract from bovine) doesn't have this
> > > > > behaviour.
> > > > > What can I do?
> > > > >
> > > > > Thanks
> > > > >
> > > > > Mariella Parisi





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