his-tagged protein purification with metal chelate columns

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Thu Jul 27 15:20:24 EST 1995


 In article <pemanuel-2707951248080001 at beaker.ab.umd.edu>
 pemanuel at umabnet.ab.edu (Peter) writes:

| I got an article out of tibs that there was a discussion on purification
| of his-tagged fusionproteins from e.coli in this newsgroup but I cannot
| find them any longer. I´m interested in getting the contributions to this
| discussion because I have problems with contamination and recovery using
| IMAC, too. If anyone has saved it would be very kind to send it.

: I too am interested in this topic.  I have had problems with this
: technique and would like to see if others have solved similar problems.
: Peter

Peter: 

The article in TIBS covers almost everything which was discussed here with a
little more info on the 21 kdal his-rich protein found copurifying with 6xhis
tagged proteins when isolated from E. coli.  I've e-mailed the past postings
to the previous person who asked for them, but I think the article is more
informative. No doubt we have not finished with this topic. I've gotten several
e-mail responses already. One interesting thought is that it may not be
possible to rid your column of such contaminants because they might be part of
the secretory mechanism of E. coli. However, so far I have not found anyone who
knows what the function of the 21 kdal contaminant is, or if the gene can be
deleted from E. coli without dire (lethal) consequences. If you can't get a
copy of my paper from the library, I can fax one to you.  I also have a more
extensive bibliography than those referenced in the TIBS article, which I can
e-mail.

@article{Hengen1995Jultibs, 
author = "P. N. Hengen",
title = "Methods and reagents - Purification of {His-Tag} fusion
proteins from {{\em Escherichia coli}}",
journal = "Trends in Biochemical Sciences", 
volume = "20", 
number = "7", 
pages = "285-286",
month = "jul", 
year = "1995"}

-Paul.

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