Binding to GST resin

Cyril Privenzentzev cprivenz at igr.fr
Mon Jan 10 12:27:20 EST 2000


Dear Netters:
I tried to purify GST-fusion protein using Glutathione Sepharose 4B
(Pharmacia). The protein is 30kDa DNA-glycosylase With high affinity to
DNA. Finally, I've got a good band. However, about 80% of activity
remains in the crude extract. Re-use of the flow-through with new batch
of resin resulted in nothing-most of the activity goes through the
resin.How can I improve the binding of my protein? High salt? More
detergent? 
Any ideas will be highly appreciated.
Sincerely,
Cyril VP

-- 
Dr. Cyril V. Privezentzev
Group "Reparation de l'ADN
CNRS UMR 8532  
Institut Gustave-Roussy
Pavillon de Recherche II
39 rue Camille Desmoulins
94805 Villejuif Cedex
FRANCE
         
Tel. +33 (0)1 42 11 54 04
Fax  +33 (0)1 42 11 52 76
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