Fitc conjugated anti human cd40

For surface staining, single cell suspensions prepared from peripheral blood, spleen, or bone marrow of humanized mice were stained with surface markers and analyzed on a CyAn ADP (Dako). For intracellular cytokine staining, cells were first stained with surface markers, and then permeabilized with cytofix/cytoperm buffer (BD Bioscience), followed by intracellular staining. FITC-conjugated anti-human CD40, CD56, PE-conjugated anti-human CD303, PE/Cy5-conjugated anti-human CD4, CD86, PE/Cy7-conjugated anti-human CD3, HLA-DR, PB-conjugated anti-human CD4, CD14, IL-2, APC-conjugated anti-human CD123, CD11c, TNF-α and APC/Cy7-conjugated anti-human CD45 were purchased from Biolegend. Pacific orange-conjugated anti-mouse CD45, PE/Texas red-conjugated anti-human CD3, CD19, CD8 and LIVE/DEAD Fixable Aqua (LD7) Dead Cell Stain Kit were purchased from Invitrogen. Data were analyzed using Summit4.3 software (Dako).