histidine-tagged proteins

millerbs at LP.MUSC.EDU millerbs at LP.MUSC.EDU
Thu Apr 8 08:01:55 EST 1993


In response to the question about histidine-tagged proteins and their
purificatoin, I would like to add that a vector system for generating
these proteins is available from Novagen (Madison, WI)(800)526-7319.
Their vector system is called pET.  I have not used their vector system,
but am familiar with their His-Bind resin which is easy to use.  This
method generates high yields. Purification of the tagged protein may be
carried out under denaturing condition (6M Guanidine HCl) and renaturation
may be attempted for functional studies.  Qiagen also markets a nickel-
chelating resin, NTA-resin, for purification of His-tagged proteins.
The method was developed by Hochuli, et al. J Chromatography (1987) 411:177.
The first utilization of this technique for protein purification was 
published in BIO/TECHNOLOGY, November, 1988, again by Hochuli, et al.
Gentz et al. used this method to purify His-Tat in PNAS (1989) 86: 821.
The reference for BIO/TECHNOLOGY paper 6:1321.  

I have used this methodology to purify large amounts of a transcription
factor which is functional after renaturation.

Good luck!

Brad Miller
MSTP Student
Dept Pharmacology
Medical University of South Carolina
Charleston, SC
millerbs at lp.musc.edu



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