Intracellular Staining by Flow Cytometry
|Detection of DC‑SCRIPT/ZNF366 in Human Mature Dendritic Cells by Flow Cytometry. Human mature dentritic cells differentated from human peripheral blood mononuclear cell derived CD14+ cells treated with 20 ng/mL Recombinant Human IL‑4 (Catalog # 204-IL) and 50 ng/mL Recombinant Human GM‑CSF (Catalog # 215-GM) for 7 days and 1 μg/mL LPS, 20 ng/mL Recombinant Human TNF‑ alpha (Catalog # 210-TA), and 20 ng/mL Recombinant Human IL‑1 beta /IL‑1F2 (Catalog # 201-LB) for last 24 hours were stained with Human DC‑SCRIPT/ZNF366 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF4707, filled histogram), or control antibody (Catalog # AB-108-C, open histogram), followed by Donkey Anti-Goat IgG (H+L) Phycoerythrin (Catalog # F0107). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.|
DC-SCRIPT (dendritic cell-specific transcript; also ZNF366) is an 85 kDa member of the Cys-Cys:His-His subfamily of zinc-finger proteins. This subfamily is principally involved in protein-DNA interactions. DC-SCRIPT appears to be DC specific, and is expressed in myeloid DC, plasmacytoid DC and Langerhans cells. Human DC-SCRIPT is 744 amino acids (aa) in length. It contains an R-K-R-K NLS (aa 77-80), a Pro-rich region (aa 110-219), eleven C2H2-type zinc-fingers (aa 255‑556) and an extended acidic region (aa 586-698). Over aa 559-744, human DC-SCRIPT is 68% aa identical to mouse DC-SCRIPT.
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