How pack Sephacryl S-100

Peter pxpst2 at unixs.cis.pitt.edu
Sat Sep 12 16:17:29 EST 1998


In article <1D81BD0F054E at serverdos.cigb.edu.cu>, juan.morales at cigb.edu.cu wrote:

> Hello netters:
> I am new, just began to worck at 24 august, my bust told me to pack a
> column of Sephacryl S-100 of 80 cm X 16 mm of diameter of gel, and the
> sample will be on 8M Urea, 10 mM beta mercaptoetanol, 50 mM Carbonate 
> bicarbonate, pH 10,6. Please, how to get:
> a) a good packed column with a number of plateu over 3000?, b) good
> resolution for a protein near 20 kDa?. Thanks in advance, Lazaro

You have to solve for your "plates" yourself utilizing known
standards(every hand packed column is a little different), as for the
column dimensions, it should be as long as tolerable(too long will
increase the longitudinal diffusion of the solute) and as narrow as
possible(to limit the latitudinal diffusion).  Sample volume size should
not exceed 5 % of column volume but preferable be less than 2%.



Peter

-- 
"Don't you eat that yellow snow
            watch out where the Huskies go"    FZ

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