Cornelius Krasel (krasel at wpxx02.toxi.uni-wuerzburg.de) wrote:
: Svend Kjaer <Svend at CAJAL.MBB.KI.SE> wrote:
: > This is a recurrent thread. However, I have the choice to tag my scFv antibody
: > fragments with either of the two tags, so which one do you recommend and
: > preferably does anyone know of Kd determinations of the M2 and 9E10 antibody
: > for their respective tags, which would eliminate the "works fine in Western
: > blot at 1:10000 for me !" kind of responses.
: Maybe it helps you to know that internal flag tags don't work. The tag has
: to be either at the N or the C terminus.
Not so. Have a look at
Howard, M., Jilling, T., Duvall, M. and Frizzell, R. A. (1996) `cAMP
regulated trafficking of epitope tagged CFTR.' Kidney International,
One of their constructs has an internal FLAG (DYKDDDDK) tag in an
extramembrane loop and it works.
Chris Boyd | from (but not \ MRC Human Genetics Unit
Christopher.Boyd at hgu.mrc.ac.uk | on behalf of) / Crewe Rd, Edinburgh
http://www.hgu.mrc.ac.uk/Users/Christopher.Boyd EH4 2XU, SCOTLAND