Ag specific Click-iT EdU?

Has anyone used the Invitogen Click-iT EdU system to measure the frequency
of Ag specific T cells?

Although upregulation of CD154 is a good marker of the total number Ag
specific T cells, the frequency of CD154+ cells in our media controls (about
1-2 per 10,000) is about the same as our frequency of Ag specific T cells. I
thought that using both CD154 and EdU might be a way to lower the noise in
the media control. I am already using LIVE/DEAD/ViViD/WTF to discriminate
dead cells...and lower noise.

My thought was to add the Ag to PBMC, do an overnight incubation, add the
EdU in the morning, allow a few hours for incorporation, and then fix.

Any experience or comments on this kind of approach?

Thanks,      Calman
_______________________
> Calman Prussin
> Laboratory of Allergic Diseases
> National Institute of Allergy and Infectious Diseases/ NIH
> Bethesda, Maryland