dry gel filtration column

Wolfgang Schechinger via methods%40net.bio.net (by novalidaddress from nurfuerspam.de)
Tue Dec 20 15:19:00 EST 2011


Hi Pepa, 

probably you just have some air trapped inside and possibly buffer salts 
have precipitated in the tubings.

The simpliest thing to try is to pump large volumes of buffer (10 to 25 
or even more column volumes of PBS with azide or just DD water/azide) at 
a low flow rate through the column (inlet at the bottom of the vertically 
mounted column!). From time to time, increase the flow rate to check if 
the performance of the column improves.

If that does not work, try to suck the buffer through with gentle vacuum. 
Lower pressure will make the trapped bubbles bigger and might get them 
out from the pores easier.

If the column may be opened without destroying it, just pour the gel into 
a beaker with buffer and remove the air by applyig vacuum in e.g. an 
exsiccator. When all the air has gone out, repack the column.

If you have a spare empty refillable column, you might consider breaking 
the selaed column, too. 

At least for sephadex based materials, this works well. Don't know about 
other stationary phases like sephacryl, however.

If, additionally, the column packing has suffered (visible "cracks"), you 
could try to make it even again by frequently reversing the flow 
direction and applying lots of pressure (flow) changes.

Or you may simply pretend that the column has been eaten by molds and 
bugs and ask your PI for a new column for your precious samples...

HTH

Wo
Am Tue, 20 Dec 2011 20:20:27 +0100 schrieb Pepa Florez Pérez:

> Hello,
> 
> I have discovered today that one of our prepacked gel filtration column
> is completely dried out. We have tried to make it work by re-hydration
> and the pressuere has decreased, but it is stil very high.
> 
> Do you know if it is possible to make it work in the same way as before?
> 
> The column seem wet now, but we cannot run it to the same flow as
> before.
> 
> Are the beads damaged somehow irreversibly when they become completely
> dried?
> 
> Thank you very nuch
> 
> Pepa



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