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We are interested in attempting to identifying Th1 and Th2 lymph node
lymphocytes of mice using intracellular cytokines by flow cytometry. Could you
provide details about how you isolated/purified lymph node T cells? Do you
have information about the proportions of cells with IFNg vs IL-4 and possibly
cells coproducing these cytokines? Has this work been published and if so
where?
Thank you for your help,
Ralph Smialowicz
U.S. EPA
RTP, NC 27711
USA
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From: Mikael Vestberg <Mikael.Vestberg@inflam.lu.se>
Subject: re:re: intracellular IL4 and IFNg
To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
Message-id: <960531.180050.25860@macpost.lu.se>
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Importance: normal
We have done analysis in mouse lymph node T cells=20
after in vitro culture. After 4 days or more of=20
culture w ConA and then a 6 hour PMA/ionomycin=20
treatment in the presence of monensin we see=20
abundant IFN gamma and some IL4. They are not=20
expressed in the same cells.
We use 50 ng/ml ConA and 50 ng/ml PMA and 1 ug/ml=20
ionomycin in the presence of 3uM Monensin.
Mikael
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