Epitope Tagging?

Jeff 'Newt' Sekelsky fznewt at peseta.ucdavis.edu
Wed Nov 1 11:38:48 EST 1995


On 31 Oct 1995, Volker Hartenstein wrote:

> Hi, > I'm Tim from the Hartenstein lab and I'm engineering an expression
> construct using epitope tagging methods.  Here's my dilemma: I've heard
> from people that the myc epitope gives very weak/no embryonic staining. 
> Has anyone used the hemaglutinin tag or the FlagTech tag or something >
else and gotten good results.


Katy Afshar from our (Scott Hawley's) lab has had good success with the 
FLAG epitope and monoclonal antibodies from Eastman Kodak.  The anti-FLAG 
initially available gave background bands on Westerns.  However, Katy 
found one, M5, that gave a strong signal with no background.  Kodak has 
now made this Ab commercially available.  The sequence recognized by the 
Ab is DYKDDDDK.

Afshar et al. (1995) DNA binding and meiotic chromosomal localization 
of the Drosophila Nod kinesin-like protein.  Cell 81:129-138.

Afshar et al. (1995) Identification of the chromosome localization domain 
of the Drosophila Nod kinesin-like protein.  J. Cell Biol. 130 (4), in press.


Jeff Sekelsky




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