rsteinbe at etowah.uokhsc.edu
Tue Jul 1 11:17:32 EST 1997
Jared Head wrote:
> I've been making home-made affinity columns by putting some glass wool at
> the bottom of a disposable 20ml syringe and pouring the sepharose in on
> top. It works, but is a bit unreliable; sometimes the glass wool allows
> resin to leak out, other times there's too much glass wool and the drip
> rate is very slow.
> Has anybody got any better methods for keeping the resin in, or can you
> buy cheap plastic columns with a filter already in the bottom (which would
> be ideal)?
> Thanks in advance,
> Jared Head at the Department of Biochemistry, University of Bristol
> "A computer lets you make more mistakes faster than any invention in human
> history - with the possible exceptions of handguns and tequila."
> Mitch Ratliffe
We routinely use circles cut from glass fiber filters (Whatman GF/C) as
supports for columns in syringe barrels. You can cut them by hand or
with a cork borer. Just push them gently to the bottom of the syringe
using the syringe plunger (first wipe off excess lubricant with a
Kimwipe)-- be careful not to crush the fibers by pushing against the
bottom-- and use a long Pasteur pipet to seat the edges of the filter in
place. After the filter is in place, fill the column with water and let
it run through to wet the filter and to be sure that nothing is clogged.
While the water is running through, tap gently and reseat the edges of
the filter with the Pasteur pipet to dislodge any trapped air bubbles.
Then stop the column (you can purchase disposable 2-way or 3-way luer
stopcocks for this purpose) and pour the resin.
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