problems of dialysis...
Dr E. Buxbaum
EB15 at le.ac.uk
Fri May 16 08:53:54 EST 1997
mange at cismsun.univ-lyon1.fr wrote:
>We are trying to purify a transcription factor on streptavidin
>magnesphere. After one passage on magnesphere, we will have to dialyse the
>proteins at 4°C and for a short period...does anyone have any suggestions
>for this last step (centricon, dialysis tubing...)?
You do not state which volume of sample you want to handle, but I suspect
it will be rather small. In such cases I had good success placing the sample
into the lids of Eppendorf vials. The conical part of the vials have to be cut
of first. You then span the dialysis membrane over the rim of the vial lid
and close the ring which remained from the vial over it. This gives you a
dialysis chamber of approx. 300 ul for a 1.5 ml vial, and about 100 ul for
the 0.5 ml vial.
Remember to boil the dialysis membrane in 100 mM NaHCO3, 10 mM EDTA before
using it to remove contaminants, in particular proteolytic enzymes. The
prepared membranes should be handled with gloves only, they can be stored
in 20% ethanol.
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