Protein purification with intein tag

OBF het_obf at rediffmail.com
Wed Oct 30 05:25:41 EST 2002


Hi everybody!
I have purified a protein with an C-terminal his-tag. However, it
seems to be inactive, presumably due to oxidation of the protein
during the Ni-affinity chromatography or due to the presence of the
His-tag (I didn't engineer in a protease recognition site, so Ican't
cleave off the His-tag).
So, I am thinking of using the intein-tag technology developed by New
England Biolabs (see: http://www.neb.com/neb/products/pfp/6900.html),
but before I pay $400 I would like to know if people here can tell me
some of their experiences with this system: does it work with all
proteins? is the cleavage of the tag 100%? is the final eluted protein
>95% pure (on the picture they show on the above website, the final
protein doesn't look completely pure)?

Thanks in advance!
Willem



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