Disposable Columns

Dima Klenchin klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Thu Jul 3 10:52:39 EST 1997


In article <33B92D9C.E75 at etowah.uokhsc.edu>, rsteinbe at etowah.uokhsc.edu wrote:
#Jared Head wrote:
#> 
#> Hi,
#> 
#> 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
#> 
#> --
#> 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.

Don't do it. Many proteins absorb to these filters nearly irreversibly. The 
filters are thick enough to have high capacity, too. 

- Dima
 



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