Intracellular Staining by Flow Cytometry
|Detection of IL‑27/IL-35 EBI3 Subunit in KG-1 Human Cell Line by Flow Cytometry. KG‑1 human acute myelogenous leukemia cell line treated with 40 ng/mL Recombinant Human IL‑18/IL‑1F4 (Catalog # B001-5) for 24 hrs was stained with Human IL‑27/IL-35 EBI3 Subunit Monoclonal Antibody (Catalog # MAB6456, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.|
IL-27 is a heterodimeric cytokine comprised of the IL-12 p35-related protein, p28, and the IL-12 p40-related protein, EBI3 (Epstein-Barr virus-induced gene 3). IL-27 is expressed by monocytes, endothelial cells and dendritic cells. It binds TCCR/WSX-1 on naive CD4+ T cells and induces the expression of a functional IL-12 receptor, making these cells sensitive to IL-12-mediated Th1 cell development. Human EBI3 also associates with the p35 subunit of IL-12 to form IL-35 which is important for immunosuppressive Treg cell induction. EBI3 is 61% amino acid (aa) identical to mouse EBI3 and includes an 20 aa signal peptide anda 209 aa mature protein with two fibronectin type III domains.
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