Freezing GSH-agarose for "pull-down fever (PDF)"

Paul pd1 at mole.bio.cam.ac.uk
Wed Mar 8 11:17:46 EST 2000



Fred wrote:

> We all know that freezing agarose or sepharose beads in aqueous
> solutions fractures them and ruins them for gel filtration.  I want know
> if they actually break apart into small pieces.  In other words, can
> they still be used for pull-downs.
>
>

We store our favourite proteins for pull-down assays on glutathione agarose
or
amylose resin at -20 °C in 20 - 50% glycerol (depending on who made the
prep).
Even at the lower glycerol concs. where the mixture obviously freezes the
beads
still work fine for pulldowns and don't seem to behave any differently to
"fresh"
stuff.  I don't think we've kept anything as long as a year (yet), but this
approach
has worked well for periods of months at least.

Paul Digard






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