Strep-tag

Dr. Arthur L. Kruckeberg bmbckh at biovax.leeds.ac.uk
Thu Dec 21 09:11:59 EST 1995


Hi

re: Strep-tag as described by Schmidt and Skerra "The random peptide
library-assisted engineering of a C-terminal affinity peptide, useful
for the detection and purification of a functional Ig Fv fragment"
Journal: Protein engineering; 1993; vol. 6(3); pp. 109-122.

I am thinking of using the strep-tag (as described in the above, read
it's a good paper) for labelling a membrane transport protein at the
C-term. The protein is a homologue of the sugar transport family.
Labelling at the C-term. would facilitate visualisation and
quantitation by western blot without the need to produce specific
anti-sera, and affinity purification using strepavidin-agarose. Has
anybody had any experience using this system for the purification of
such proteins and has anybody had any experience using this system in
the presence of detergents? If so, could they advise me of the
pitfalls, expected yields and level of purity? I've also looked into a
few suppliers of relevant reagents esp. streptavidin-agarose; could
anyone advise me as to how effective this agarose is for purification
from different suppliers and its repeated use?
Thanks for the advice in advance.
Merry Christmas and a Happy New Year.
Chris
------------------------------------------------------
Chris Hoyle

   Department of Biochemistry and Molecular Biology
University of Leeds             phone +44 0113 2333172
Leeds LS2 9JT                   FAX   +44 0113 2333167
Great Britain           e-mail bmbckh at biovax.leeds.ac.uk
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