glycerol removal by dialysis - protocol and pitfalls? (Jan/29/2010 )

Hi,
I need to remove glycerol from my stock antibody solution (PBS with 50% glycerol). Anyone have any experience doing this? I was planning on diluting my sample 1:5 and then dialyzing against 100volumes excess PBS for an hour or so, do you think this will work as intended? Thanks in advance!

PKNubbins on Jan 29 2010, 10:08 PM said:

Dialysis should work fine. However it is often forgotten that it is a very slow process. I would suggest using ca 50 volumes of PBS for 36 hours. During this time replace the buffer 3 times.

Your antibody solution will increase in volume during dialysis so remember to use extra tubing


Hope this helps

Antibody novice, but could you dilute the Ab/glycerol solution, load onto anafinity column and elute in a smaller volume of your desired buffer?

klinmed on Jan 30 2010, 04:34 AM said:

if you make the bag tight then you can avoid a significant increase in volume.

mdfenko on Feb 2 2010, 05:55 PM said:

But also risk rupture of the dialysis tubing!

klinmed on Feb 3 2010, 02:40 PM said:

never happened (to me or anyone with whom i worked). the tight bag will resist the increase in volume and is strong enough to not burst. if you had any burst then it was probably damaged during loading or defective.

also, if the sample is low volume then the tubing for low volume is usually thicker walled and even more resistant to bursting.