Elution from Glutath. beads

ossipow ossipow at sc2a.unige.ch
Mon Mar 13 13:41:05 EST 1995


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 ??
  By the way I have another question : I run rat liver nuclear extracts
over GST-fusion column to "fish out" proteins that would interact with
the factor I am working with. As a control I use a column with GST alone;
I often have IRREPRODUCIBLE and (presumably & hopefully) unspecific 
binding on the GST alone column. What would you recommend to avoid those
 problems ?

  Thank you in advance
   Vincent Ossipow



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