IHC with anti-FLAG Ab - (Jan/08/2010 )

Hi,

I need to do an IHC on frozen tissue (skin) and detect a FLAG-tagged nuclear protein. I have tried Sigma M2 and F7425, no staining... Unfortunately, I don't have a positive control. Western does not work, too few cells (2000/mouse).
Does anyone know a good antibody or protocol? How about antigen retrieval, would that be better?
Any help/suggestions really appreciated!

Cheers,
Minna

"FLAG" is a trademark belonging to sigma, so no other company should be able to sell an antibody with that name. Other companies sell their own version of the antibody recognizing the "FLAG" peptide: DYKDDDDK

Try doing some parallel experiments (Western and immunocytochemistry) with some cultured cells transfected with a plasmid expressing your protein. At least then you can be sure your reagents are working.