IL-18/IL-1F4 Antibody (925008) [Unconjugated]

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.

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.

Recombinant Human IL‑18/IL‑1F4 protein was serially diluted 2-fold and captured by Rabbit Anti-Human IL‑18/IL‑1F4 Monoclonal Antibody (Catalog # MAB91243) coated on a Clear Polystyrene Microplate (Catalog # 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 (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

• Reactivity: Hu
• Applications: ELISA, CyTOF-ready, Flow, ICC/IF
• Clone: 925008
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

IL-18/IL-1F4 Antibody (925008) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant Rhesus monkey IL-18/IL-1F4
  Tyr37-Asp193
  Accession # Q14116
• Specificity: Detects Rhesus monkey and Human IL-18/IL-1F4 in direct ELISAs.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • ELISA
  • Immunocytochemistry 5-25 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for IL-18/IL-1F4 Antibody (925008) [Unconjugated]

• Iboctadekin
• IFN-gamma-inducing factor
• IGIF
• IGIFIL-1 gamma
• IL18
• IL-18
• IL-18MGC12320
• IL-1F4
• IL1F4iboctadekin
• IL-1g
• Interferon gamma-inducing factor
• interleukin 18 (interferon-gamma-inducing factor)
• Interleukin-1 gamma
• interleukin-18

Background

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).

1. Arend, W.P. et al. (2008) Immunol. Rev. 223:20.
2. Giavedoni, L.D. et al. (2001) J. Interferon Cytokine Res. 21:173.
3. Ghayur, T. et al. (1997) Nature 386:619.
4. Gu, Y. et al. (1997) Science 275:206.
5. Boraschi, D. and C.A. Dinarello (2006) Eur. Cytokine Netw. 17:224.
6. Novick, D. et al. (1999) Immunity 10:127.
7. Torigoe, K. et al. (1997) J. Biol. Chem. 272:25737.
8. Born, T.L. et al. (1998) J. Biol. Chem. 273:29445.
9. Bufler, P. et al. (2002) Proc. Natl. Acad. Sci. USA 99:13723.
10. Takeda, K. et al. (1998) Immunity 8:383.
11. Park, S. et al. (2007) Cell. Mol. Immunol. 4:329.
12. Yoshimoto, T. et al. (1998) J. Immunol. 161:3400.
13. Hoshino, T. et al. (2001) J. Immunol. 166:7014.
14. Iannello, A. et al. (2009) AIDS Rev. 11:115.
15. Rabkin, S.W. (2009) Nat. Clin. Pract. Cardiovasc. Med. 6:192.
16. Netea, M.G. et al. (2006) Nat. Med. 12:650.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for IL-18/IL-1F4 Antibody (MAB2548)(2)

Publications using MAB2548

• Linqiang Huang, Mengting Liu, Wenqiang Jiang, Hongguang Ding, Yongli Han, Miaoyun Wen et al. Bradykinin/bradykinin 1 receptor promotes brain microvascular endothelial cell permeability and proinflammatory cytokine release by downregulating Wnt3a Journal of Biochemical and Molecular Toxicology 2022-12-01 [PMID: 36111657]
• Linqiang Huang, Mengting Liu, Wenqiang Jiang, Hongguang Ding, Yongli Han, Miaoyun Wen et al. Bradykinin/bradykinin 1 receptor promotes brain microvascular endothelial cell permeability and proinflammatory cytokine release by downregulating Wnt3a Journal of Biochemical and Molecular Toxicology 2022-12-01 [PMID: 36111657] (Immunocytochemistry, Human)

Bioinformatics

• Uniprot:
  • Q14116()