Monensin

Mr. James Freeman; (jfreeman@umbc.edu)
Fri, 1 Sep 1995 12:17:03 -0400 (EDT)

Vivian,

You should try to get your hands on the article in _J. Immunol. Meth._ by
Jung et.al. (159 (1993) 197 - 207) entitled "detection of intracellular
cytokines by flow cytometry." I have used monensin at the concentration
of 1 uM for the detection of intracellular IL-4 and g-IFN. With this
concentration and an incubation period of 2 - 6 hrs (either on ice or at
37oC), I have observed good resolution of intracellular cytokines. The
good news for you is that I have also seen good staining of surface
proteins after treatment with monensin and saponin.

About the subject of solublity, I had no luck with DMSO, went ahead and
dissolved it in 100% EtOH. I got the results mentioned above. The cells I
used were freshly isolated splenocytes and sarcoma cells. The surface
markers remained intact.

I hope this helps. Good luck!

Jim Freeman
Dept. of Biological Sciences
University of Maryland, Baltimore County
5401 Wilkens Avenue
Catonsville, Maryland 21228

E-mail: jfreeman@umbc.edu