Human IL-18/IL-1F4 Antibody Summary
Accession # Q14116
This antibody functions as an ELISA detection antibody when paired with Rabbit Anti-Human IL‑18/IL‑1F4 Monoclonal Antibody (Catalog # MAB91243).
This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human Total IL-18 DuoSet ELISA Kit (Catalog # DY318-05) for convenient development of a sandwich ELISA or the Human Total IL-18/IL-1F4 Quantikine ELISA Kit (Catalog # DL180) for a complete optimized ELISA.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of IL‑18/IL‑1F4 in Human PBMC's. IL-18/IL-1F4 was detected in immersion fixed human PBMC’s using Mouse Anti-Human IL-18/IL-1F4 Monoclonal Antibody (Catalog # MAB2548) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of IL‑18/IL‑1F4 in Human THP-1 cell line by Flow Cytometry. THP-1 Human acute monocytic leukemia cell line was stained with Mouse Anti-Human IL-18/IL-1F4 Monoclonal Antibody (Catalog # MAB2548, filled histogram) or isotype control antibody (MAB002, open histogram), followed by PE-conjugated Anti-Mouse IgG Secondary Antibody (F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin. View our protocol for Staining Intracellular Molecules.
Human IL‑18/IL‑1F4 ELISA Standard Curve. Recombinant Human IL-18/IL-1F4 protein was serially diluted 2-fold and captured by Rabbit Anti-Human IL-18/IL-1F4 Monoclonal Antibody (MAB91243) coated on a Clear Polystyrene Microplate (DY990). Mouse Anti-Human IL-18/IL-1F4 Monoclonal Antibody (Catalog # MAB2548) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998) followed by Substrate Solution (DY999) and stopping the enzymatic reaction with Stop Solution (DY994).
Preparation and Storage
- 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.
Interleukin-18 (IL-18), also known as IL-1F4 and IFN-gamma inducing factor (IGIF), is a member of the IL-1 family of cytokines and is a key molecule in the innate immune response (1). Rhesus IL-18 is synthesized as a 24 kDa proprotein that contains a 36 amino acid (aa) propeptide and a 157 aa mature region (2). Under inflammatory conditions, the propeptide is cleaved by Caspase-1 in the cytoplasm to liberate the mature nonglycosylated 18 kDa monomeric IL-18 (3, 4). Mature rhesus IL-18 shares 96% aa sequence identity with human IL-18 and 60-76% with mouse, rat, canine, feline, and porcine IL-18. IL-18 is secreted by a variety of cell types including macrophages, dendritic cells, and epithelial cells (1, 5). Circulating mature IL-18 is sequestered by soluble IL-18 binding proteins (IL-18 BP) that inhibit IL-18 bioactivity (6). IL-18 interacts with the widely expressed IL-18 R alpha which then recruits the signaling subunit IL-18 R beta (7, 8). The IL-1 family member IL-1F7 also binds to IL-18 R alpha but does not recruit IL-18 R beta or induce signaling (9). IL-1F7 binds IL-18 BP and enhances its neutralizing effect on IL-18 activity (9). IL-18 synergizes with other cytokines to activate NK, Th1, and Th17 cells and to increase the production of IFN-gamma (1, 5, 10-12). IL-18 can also promote Th2 cytokine release which reduces the effectiveness of antiviral responses (13, 14). Increased levels of active IL-18 contribute to the severity of autoimmunity and hypertension, while deficiency of IL-18 results in symptoms of metabolic syndrome (1, 5, 15, 16). In cancer, IL-18 stimulates Th1 and NK cells to target tumor cells, but it can also promote angiogenesis, metastasis, and tumor cell immune evasion (11).
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