gelfiltration on superdex 75 pg

Randy randall.cameron2 at gte.net
Mon Oct 16 13:06:29 EST 2000


Sigrid Van Boxstael <svboxsta at vub.ac.be> wrote in message
news:39D36691.D21C4C49 at vub.ac.be...
> Hi,
>
> I'm trying to purify my protein.
> I bring 10 mg (already 90 % pure) on a Pharmacia superdex 75 pg column
> and
> I can only recuperate 1.5 mg in the obtained peak.
> The buffer I used : Tris 20 mM pH 8.2, 2 mM mercapto-ethanol, 150 mM
> NaCl.
> When I clean the column after the run there is coming protein of it.
> I presume that my protein sticks on the column.
>
> I think that increasing the salt concentrations might  be the solution.
> Does anybody know either this might help and if yes until what
> concentration
> I can increase it?
>
>
> Thanks,
> Sigrid

I had the same problem also when working with a glucosidase, it also bound
to a Toyopearl resin only not as strongly.  I had to go to a Biogel P-100
column.







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