Sephacryl 500 column problem...
barry.ford at drdc-rddc.gc.ca
Mon Nov 4 15:41:58 EST 2002
In article <aq5pib$64f$07$1 at news.t-online.com>, engelbert_buxbaum at web.de
> Szymon wrote:
> > I once wanted to separate some protein aggregates. I mean, big ones from
> > smaller and from monomeric protein. Through I've been strongly
Using the Sephacryl resins usually means you have a fibre "filter" at
each end, which is nominally to prevent resin loss through the frits.
Unfortunately, if you have an early extract, with some DNA or RNA in it
(likely), the fibre filter entrains the nulceic acids very nicely.
This is the same principal as those glass fibre extraction resins.
Change the filter, and pressure will drop. Even when you refresh the
column resin, if you reuse the fibre disc, the damned thing goes to high
Be prepared to change the fibre disc often, maybe every other run, to
maintain consistent results.
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