T cell depletion
Brian Rabinovich (brian.rabinovich@utoronto.ca)
Sun, 7 Jul 1996 19:22:26 -0400
I have developed a simple assay for assessing the intracellular cytokine
expression of IL-10 (PE-Caltag) and TNF-alpha (FITC-Pharmingen) in
stimulated monocytes (Tricolor CD33-Caltag) cultured in non-adherent costar
24 well plates (PMA/Iono/BFA for 4 hrs in OPTI-MEM/antibiotics/10%FCS/1X B
mercaptoethanol). I am stimulating PBMC isolated using ficoll from
heparinized human venous blood.I would like to deplete the stimulated PBMC
of T cells after stimulation using anti-CD3 mAbs and complement. Does
anyone have a simple protocol for doing so (i.e. concentrations of mAb and
C'/# of cells or ml). I am looking at approximately 1-2 million PBMC per
well. Any help would be appreciated. Thanx
Brian Rabinovich
Research Institute
The Wellesley Hospital
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