epitope tagging

at unixg.ubc.ca at unixg.ubc.ca
Fri Sep 26 18:23:26 EST 1997


Dear Colleagues,

I am looking for any information people might have about using epitope tags in
plants.

I want to eitope tag a presumably ER membrane bound protein of A. thaliana and
study its localization by TEM (immunogold labeling). I have been thinking of
trying either the c-Myc or the hemagglutinin epitope.

If you have any experience in tagging plant proteins I would appreciate to
hear about it. I am especially interested in the following questions:
- Will either of the two epitopes be detectable by immunogold labeling or is
  there a high risk of them being hidden (e. g. by the fixation method)?
- Do you know anything about the crossreactivity of either of these
  commercially avalaible antibodies to host proteins?
- How many epitopes do you add for getting a specific and strong signal?
- How important is a knowledge of the protein's structure in determing where
  to add the epitope efficiently? How successful have people been in just
  attaching the epitope to one of the ends of the protein?

Thanks a lot for your help!

Maren Rossak (rossak at botany.ubc.ca)






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