Hello,
I am working with an E.coli polytopic membrane protein that is expressed
at a VERY low level (probably around 100-500 molecules per cell) and I
have a lot of trouble visualising it on SDS-PAGE. I have a rabbit
polyclonal antibody raised against a domain of this protein that is
highly conserved in many homologues. This leads to the detection of
these homologues in E.coli extracts on Western blots, thereby rendering
the antibody of little use for immunoprecipitations of specific
complexes with my protein. It is not yet possible to raise polyclonal
sera against non-conserved domains.
So, I thought that epitope tagging the protein might help with specific
detection. I have already His tagged it and used a His tag monoclonal
antibody but the detection is very poor (probably since its a
monoclonal). I wonder if anyone out there has had similar experiences
and/or can suggest other epitope tags - preferably small and with a
polyclonal antibody raised against it.
Any help would be much appreciated.
Replies by e-mail to : blight at igmors.u-psud.fr
Thanks.
Mark
____________________________
Dr. Mark A. Blight,
Institut de Génétique et Microbiologie,
Bâtiment 409,
Université de Paris XI,
91405 Orsay cedex,
France.
Tel: +33 1 69416699
Fax: +33 1 69417808
e-mail: blight at igmors.u-psud.fr
____________________________
