I have extensive experience with cytokine flow in the human system with some in the mouse.
First of all, what mAbs are you using? Choice of mAbs is critical. Recently, Pharmingen has
begun directly labelling anti-cytokine mAbs in the mouse system. Try PE as a fluorochrome for
best S/N.
As far as stimulus, try PMA/ionomycin. For cytokine flow it always works better than mitogen.
Once you have that working, you can change to a less potent stimulus. Check out the Nov '95
JEM for a paper by O'Garra and Oppenshaw-it shows some of the best mouse staining in print.
Best of luck,
Calman Prussin
NIAID/ NIH
