GST fusion protein won't elute

T. S. Pillay tpillay at ucsd.edu
Sat Nov 12 19:19:36 EST 1994


Hi, Netters

My colleague working alongside me has this problem:
He has tried to purify the GRB2 SH2 domain but he finds that the protein
does not elute off the beads with glutathione(GSH).  I know this sounds
weird but has anyone experienced a similar problem with this or any other
fusion protein.  He gets good induction of expression when looking at
whole cell lysates but very poor yields with purification.  When he 
takes the GSH-sepharose beads after elution with GSH and boils it in
laemlli buffer- he finds "tons" of the fusion protein indicating that the
fusion protein is sticking avidly to the beads and not eluting off.
 Any ideas/suggestions would be gladly appreciated.



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