Hello, Paul,
You want to purify His/GST-tagged protein.
(if tags are separeted into N and C terminus,better to purify :-)
L-Histidine also works as the eluent.
(but I don`t know whether it also works well to purify GST-tag )
I found this information shown below, you can also search easily
(here also shown L-his works better than EDTA.)
http://tto.biomednet.com/
As alternative way to remove imidazole from 1st eluted sample,
how about using buffer exchange by dialysis ???
And after that, try to purify, if needed.
In our lab, my colleage constructed
NH2- GST - [ORF] -HA-His-COOH.
In this case, he could purify enough only by Ni-NTA.
Hope to be your help,
Take
***********************************************************
Takeshi FUKUHARA
Laboratory of Cell and Developmental Biology (c/o Dr.Takeo Kishimoto),
Department of Biological Sciences,
Graduate School of Biosciences and Biotechnology,
Tokyo Institute of Technology.
B2-833 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan
Phone : 81-45-924-5725/Fax : 81-45-924-5738
e-mail: tfukuhar@bio.titech.ac.jp
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