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Respnse to His-tagging of Plant Expressed proteins

Thomas Leustek leustek at AESOP.RUTGERS.EDU
Wed Jul 26 13:45:10 EST 2000

<x-flowed>I received a very poor response to my question about the 
use of the His-tag
to immunolocalize proteins expressed in plants. The only response was from
Shaul Yalovsky, which sounds as if he had problems with the antibodies (see
message that follows). I have tried the Quiagen anti-His antibody (part
number 34698) on Western blot and could not detect background or
cross-reactivity with any native Arabidopsis or Brassica protein, although
the antibody efficiently detects a His-tagged Arabidopsis protein
(expressed in E.coli).

I think someone needs to give His-tagging in plants a serious try.

Tom Leustek

At 08:41 AM 7/25/00 +0200, you wrote:
>His tagging may work for purification but is usually less suitable for
>immunocytological purposes because of high background. An option is to
>combine the His tag with a mAb tag such as (His)6-HA.   If you will decide
>to use His-tag alone try to find out about different makers as the quality
>of Abs can differ quiet substantially.  If you do decide to use a mAb tag
>as well I would recommend to avoid the "Flag tag" since we have had very
>bad experience with it.
>Good luck.
>Shaul Yalovsky
>Department of Plant Sciences
>Britania Bldg.
>Tel Aviv University
>Ramat Aviv, Tel Aviv 69978
>Phone: 972-3-640-5268
>Fax:      972-3-640-9380

Thomas Leustek
Associate Professor
Rutgers University
Biotechnology Center for Agriculture and the Environment
59 Dudley Road
New Brunswick, New Jersey 08901-8520
Telephone: (732)932-8165, extension 326
Fax: (732)932-0312
email: leustek at aesop.rutgers.edu



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