HIS6 Tagged Int. Membrane Prot?

U12201 at uicvm.uic.edu U12201 at uicvm.uic.edu
Fri Mar 17 18:15:26 EST 1995


I have used the IDA from Pierce with non-ionic detergent - it
worked fine - also IN THEORY it should work fine, so my guess
is that it will be OK.
In article <rmiller-1403950102000001 at bhall01-sl.cc.emory.edu>, rmiller at bimcore.emory.edu (Richard Miller) says:
>
>   Has anyone tried to HIS6 tag an outer mitochondrial membrane protein
>for purification by IDA chromatography?  I know that the mitochondrial
>targeting sequence for my protein resides in the C-Terminus and that
>manipulations of the N-Terminus do not affect targeting to the OMM.  I'm
>using a yeast expression system which works great, but purification on a
>large scale is proving to be a real problem. I'd like to attach a His6 tag
>to the N-terminus to provide a means of easy purification, but I don't
>know if the presence of detergents (VERY low concentration 0.05% Triton)
>will affect binding of the protein to the IDA column.  If anyone has
>attempted this (for any membrane protein) with either success or failure
>please let me know.
>
>Thanks in Advance,
>
>Richard Miller
>Department of Biochemistry
>Emory University
>Atlanta, GA



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