Elution from Glutath. beads

Zal Suldan Z-Suldan at ski.mskcc.org
Tue Mar 14 08:44:05 EST 1995

In article <1995Mar13.184105.2199 at news.unige.ch>
ossipow at sc2a.unige.ch (ossipow) writes:

> I have troubles and irreproducibility when I try to elute GST fusion
> proteins from glutathione sepharose beads (purchased from Pharmacia),
> even with a new batch of reduced glutathione (therefore presumably 
> not oxidized) that I use at 10 to 15 mM in the presence of 0.1% triton 
> for 10 min. at 37c. Does anyone has a good trick ??

This is strange. We get great, reliable elution at 10minutes, Room
Temp, 5.0mM glutathione, and no Triton. And the Sepharose is even
better than agarose.

Have you checked the pH of the glutathione solution used to elute? I've
found elution drops off significantly when pH is below 7.5 with best
elution when pH = 8.0 - 9.0

All opinions are mine and not of MSKCC

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