HELP: Purification of his-tagged protein

Christophe Andreoli andreoli at cell.biol.ethz.ch
Wed Nov 27 04:04:39 EST 1996


  Dear David,



   I have expressed the last year some his-tagged proteins.
Some of my idea:
When the protein is too low expressed (it seems to be true for you, did you
make a western blot, do you see signs of degradations?), it is rather
difficult to clean the protein from the Ecoli contaminants. In my hands,
the GST system is rather better (very little cross purification).
However, I found that addition of a low concentration of imidazole in the
lysis buffer improves very good the thing, and leaves behind most of the
contaminants.
 I hope it helps. Contact me if you want more informations

    Christophe Andreoli

----------------------------------------------
Christophe Andreoli, Institute for Cell Biology
HPM F 46, ETH H=F6nggerberg
CH-8093 Zurich, Switzerland
phone: ++41 1 633 33 93, FAX: ++41 1 633 10 69
email: andreoli at cell.biol.ethz.ch=20





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