In article <954pcp$f4s$1 at uranium.btinternet.com>,
"Richard Buick" <Richard.Buick at btinternet.com> wrote:
> Hello
> I've been expressing His-tagged proteins in E. coli Origami cells, about 10
> different proteins in all. I was using a His-probe for detection and was
> getting only about half of the proteins expressed. Until I turned to
> specific antibodies for detection eg, anti-prolactin and suddenly all
> proteins were detected. All had been sequenced, the His-tag is 5` to the
> protein sequence. Does anyone know why some are detected and others not by
> the His-probe? This is a real problem because I had planned on using the
> His-tag for purification.
>
This is just a guess, but maybe they are initiating translation
downstream from the His tag site.
Nick
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Nick Theodorakis
nicholas_theodorakis at urmc.rochester.edu
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