6xHis vs. 8xHis in Dictyostelium

Kurt Thorn thorn at itsa.ucsf.edu
Mon Aug 10 15:37:18 EST 1998


I am trying to purify a 380 kDa recombinant protein from Dictyostelium for
crystallography.  I have been using a 6xHis tagged protein and have been
having a lot of trouble with poor binding capacities of the metal resin and
lots of contamination of the eluted product.  
Lately I have heard that 8x or 10x His tags work better than 6x His tags
for organisms with lots of his-rich proteins (like Dicty).  Can anybody
give me references or personal experiences of working with these larger
tags?  Any other tips on difficult his-tag purifications are welcome.


      Kurt Thorn - thorn at itsa.ucsf.edu - http://motorhead.ucsf.edu/~thorn

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