Detection of IL‑18/IL‑1F4 in Human PBMCs by Flow Cytometry.
Human peripheral blood mononuclear cells (PBMCs) either untreated (upper panel) or treated with LPS for 24 hours (lower panel) were stained with Mouse Anti-Human IL‑18/IL‑1F4 Propeptide Monoclonal Antibody (Catalog # MAB646, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin. View our protocol for Staining Intracellular Molecules.
Detection of IL‑18/IL‑1F4 Propeptide in Human THP-1 cells by Flow Cytometry.
THP‑1 human acute monocytic leukemia cells were stained with Mouse Anti-Human IL‑18/IL‑1F4 Propeptide Monoclonal Antibody (Catalog # MAB646, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram) followed by PE-conjugated anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Pro-IL-18 (pro-Interleukin 18; also pro-IGIF and pro-IL-1 gamma ) is a 24 kDa member of the IL-1 family of molecules. It is widely expressed, being produced by keratinocytes, intestinal epithelium, T cells, macrophages and osteoblasts. Human Pro-IL-18 is 193 amino acids (aa) in length. Although mature IL-18 induces IFN-gamma secretion by NK and T cells, Pro-IL-18 appears to have little intrinsic activity. Generally, active IL-18 is considered to arise from caspase-1 cleavage of Pro-IL-18 between Asp36-Tyr37. This generates an 18 kDa mature C-terminal fragment, and a 4 kDa (predicted) N-terminal prosegment that runs at 6 kDa in SDS-PAGE. Other proteases are known to process Pro-IL-18. Caspase-3 cleavage after Asp68 generates an inactive 14 kDa mature segment, Merpin beta -subunit cleavage after Asn52 generates a marginally active 17 kDa mature segment, while parasite Cys protease cleavage after Val47 generates an inactive 17 kDa mature molecule. One splice variant shows a deletion of aa 27-30. Over aa 2-36, human Pro-IL-18 shares 63% aa identity with mouse Pro-IL-18.
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