Re: INF g and TNF b intracellular staining

From: maciej simm (simm@cd4cd8.com)
Date: Fri Apr 12 2002 - 15:52:11 EST


I've tried to induce inf-g with frozen cells in the past, I find that
although the production occurs, the external immunophenotyping isn't optimal
for several molecules. CD4 seems to suffer from the thawing, as does CD56.
CD8 and 3 don't seem to change as much, neither do CD19/20.

I was also able to detect CD69 production on the same cells but only with
the strongest activators.

Also, even my most positive controls yield lower production of CD69 and
inf-g on frozen cells than their non-frozen controls.

Frozen are more likely to have higher background. But inf-g signal is
usually in the 3rd log (depending on your fluorochrome, I would recommend BD
PE or Caltag RPE conjugates)

Maciej



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