epitope tags for mammalian expression

Rupert Yip yip at rascal.med.harvard.edu
Wed Aug 20 16:21:36 EST 1997


Michael Gerdes wrote:

> Looking for some experiences on some of the epitope tags that people
> are
> using in mammalian expression systems.  Merits/cons of HA, myc, etc.
> I
> need to localize a protein (suspected membrane protein) and would like
> to
> detect by immunofluorescence and western of fractionated cells.  I
> would
> like to clone the tag into the carboxy terminus.
>
> Thanks,
>
> Michael

   In my experience, the HA is not as good as the myc because althought
the HA epitope is small and convienient, it is not very antigenic and
multiple HA epitopes are more effective (ie 2 consecutive HA epitopes is
10X better than 1 HA epitope).  In addition, the HA monoclonal antibody
from Bohringer Mannhein is not so good.  I've heard of another supplier
for HA antibody (polyclonal) but haven't tested it.  Also, you probably
know that polyclonals are better for immunoprecipitations rather than
monoclonals.

As for myc, there is more comercial sources of anti-myc so you should
get a better choice as to which antibody is better.  Also, there are
commercially available cloning vectors which contain myc as the tag but
I haven't seen any for HA.




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