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I have not compared a large number of cytokines, but for huIL-2 and IFN I
do not see a significant difference in the mean fluorescence intensity or
% positives between BFA and monensin.
The main reason for using monensin in the past was that it was dramatically
cheaper. Sigma now sells BFA at a reasonable price. It is worth having a
through discussion of the relative merits of each, as I am eager to switch
to a better reagent. I look forward to hearing from you to substantiate
your statement.
Intracellularly Backed Up in Bethesda,
Calman Prussin
Allergic Diseases Section
NIAID/ NIH
calman@nih.gov
------------------------ Forwarded Message -----------------------
Jonni S. Moore wrote:
In response to your questions concerning intracellular cytokine staining:
1. Brefeldin A is MUCH better than monensin. It is less toxic and
exerts its effects in a shorter time frame--we add it to cultures typically
4 hours before staining.
2. Pharmingen is marketing a line of anti-cytokine antibodies that they
claim are optimized for intracellular staining. We have used their
FITC-anti-mouse interferon gamma and PE-anti-mouse IL4 with success.
Several other companies have labelled anti-cytokine antibodies, but they
may not be suitable for intracellular use. You'll just have to try them.
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