Human/Rhesus Macaque IL-18/IL-1F4 Antibody Summary
Accession # AAK13416
This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human IL‑18/IL‑1F4 Monoclonal Antibody (Catalog # MAB91242).
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.
IL‑18/IL‑1F4 in Human PBMCs. IL‑18/IL‑1F4 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human/Rhesus Macaque IL‑18/IL‑1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (green). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
IFN‑ gamma Secretion Induced by IL‑18/IL‑1F4 and Neutralization by Primate IL‑18/IL‑1F4 Antibody. In the presence of Recombinant Human TNF‑ alpha (20 ng/mL, Catalog # 210-TA), Recombinant Rhesus Macaque IL‑18/IL‑1F4 (Catalog # 2548-RM) stimulates IFN‑ gamma secretion in the KG‑1 human acute myelogenous leukemia cell line in a dose-dependent manner (orange line), as measured by the Human IFN‑ gamma Quantikine ELISA Kit (Catalog # DIF50). Under these conditions, IFN‑ gamma secretion elicited by Recombinant Rhesus Macaque IL‑18/IL‑1F4 (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human/Rhesus Macaque IL‑18/IL‑1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548). The ND50 is typically 0.4-1.2 µg/mL.
Western Blot Shows Human IL‑18/IL‑1F4 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and IL-18/IL-1F4 knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Rhesus Macaque IL‑18/IL‑1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for IL‑18/IL‑1F4 at approximately 25 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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).
- Arend, W.P. et al. (2008) Immunol. Rev. 223:20.
- Giavedoni, L.D. et al. (2001) J. Interferon Cytokine Res. 21:173.
- Ghayur, T. et al. (1997) Nature 386:619.
- Gu, Y. et al. (1997) Science 275:206.
- Boraschi, D. and C.A. Dinarello (2006) Eur. Cytokine Netw. 17:224.
- Novick, D. et al. (1999) Immunity 10:127.
- Torigoe, K. et al. (1997) J. Biol. Chem. 272:25737.
- Born, T.L. et al. (1998) J. Biol. Chem. 273:29445.
- Bufler, P. et al. (2002) Proc. Natl. Acad. Sci. USA 99:13723.
- Takeda, K. et al. (1998) Immunity 8:383.
- Park, S. et al. (2007) Cell. Mol. Immunol. 4:329.
- Yoshimoto, T. et al. (1998) J. Immunol. 161:3400.
- Hoshino, T. et al. (2001) J. Immunol. 166:7014.
- Iannello, A. et al. (2009) AIDS Rev. 11:115.
- Rabkin, S.W. (2009) Nat. Clin. Pract. Cardiovasc. Med. 6:192.
- Netea, M.G. et al. (2006) Nat. Med. 12:650.
No product specific FAQs exist for this product, however you mayView all Antibody FAQs
Cell and Tissue Staining Kits
Reviews for Human/Rhesus Macaque IL-18/IL-1F4 Antibody
There are currently no reviews for this product. Be the first to review Human/Rhesus Macaque IL-18/IL-1F4 Antibody and earn rewards!
Have you used Human/Rhesus Macaque IL-18/IL-1F4 Antibody?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image