Flow cytometry assay for intracellular cytokine levels of immune cells includes fixing and permeabilizing the cells with PermeaFix (TM) reagent

摘要

Method for determining the intracellular cytokine levels of immune cells in body fluid, organ or tissue samples comprises: (a) isolating mononuclear cells; (b) restimulating the mononuclear cells in vitro; (c) fixing and permeabilizing the cells with PermeaFix (TM) reagent; (d) staining cell-surface antigens and intracellular cytokines; and (e) analyzing the cells by flow cytometry. Method for determining the intracellular cytokine levels of immune cells in body fluid, organ or tissue samples comprises: (a) isolating mononuclear cells by density gradient centrifugation; (b) restimulating the mononuclear cells in vitro; (c) fixing and permeabilizing the cells with PermeaFix (TM) reagent; (d) staining cell-surface antigens and intracellular cytokines with dye-labeled monoclonal antibodies; and (e) analyzing the cells by flow cytometry.