Epitope tags: Methionines?

Arle Kruckeberg kruckeba at biovx1.biology.ucla.edu
Wed Aug 14 12:18:37 EST 1996


In article <Pine.GSO.3.95.960807113817.7726B-100000 at chip.ucdavis.edu>
Steven Theg <fztheg at peseta.ucdavis.edu> writes:

> Is anyone familiar enough with epitopes to know whether any for which
> commercial antibodies are available contain methionines?  I would like to
> epitope tag a protein that we translate in vitro.  The protien has no
> methionines for [35S] labeling, so use presently use [3H]leucine.  I'd
> like to introduce methionines into this protein at the same time I add the
> epitope tag.  So, it would be nice if the epitope containined methionines,
> the more the better, but two is probably the minimum.

Hi Steve--

The bacteriophage T7 gene 10 epitope is

M A S M T G G Q Q M G

for a total of 3 Mets. It is from the N-terminus of the gene 10
protein,
but is recognized fine when tagged to the C-terminus of my protein.
The antibody is available from Novagen, and does not cross-react with
any other bands in yeast crude extracts when I do a Western. Note that
the epitope has a net charge of zero.

I have no affiliation with Novagen!
Arle
*************************************************
a.k.a. Dr. Arthur L. Kruckeberg
 Department of Biological Chemistry
UCLA School of Medicine
Los Angeles CA 90024
(310) 825-8363     FAX (310) 206-5272
e-mail kruckeba at biovx1.biology.ucla.edu
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