Re: Intracellular staining of cytokines

Andreas Schroeder (h82@ix.urz.uni-heidelberg.de)
Sat, 7 Dec 1996 11:23:49 +0100

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>Dear flow(ers)!
>We are struggeling with intracellular staining of INFg/IL4 using BD
>FASTimmune cytokine system. The cells seems to die and there are not
>many cells picking up the cytokine stain. Using other markers for
>intracellular staining in the same system seems to function (activation
>of cells with PMA-Ionom.,adding Brefeldin A and looking for
>intracellular CD69). Other lab's do also have problems when following
>the procedure given by BD.
>
>If anyone has succeded -what is the trick? What is a good positive
>control for a-IL4 and a-INFg?

Sorry,

I don't have any answers but further questions to this subject:

Has anyone here tried Pharmingens reagents for detection of intracellular
cytokines? Anyone who has suceeded in detecting intracellular IFN-g/IL-4
following stimulation by crosslinking receptors (i.e. CD3xCD28)?

Any answers are greatly appreciated.

--
Dr. med. Andreas Schroeder
Institute of Immunology
University of Heidelberg
Germany

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