Does anybody have experience in intracellular cytokine or NFkB staining for FACS in human neutrophils?

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Sylvia Samaniego

I've done cytokine staining in primary murine neutrophils. For cytokine staining I pre-incubate with GolgiPlug and then used BD Cytofix/perm (Saponin based) with good results.

Can you be more specific about what you've tried and where it's getting tricky? Have you had success with the same antibody in other cell types? Have the neutrophils been activated?

Christopher Lee Fogarty

Thanks for the info Sylvia. Yah, they are activated cells and there has been no problem in other cell populations for either cytokines or NFkB. I got some suggestions from the cytometry mailing list that I am going to try out.