Human/Rhesus Macaque IL-18/IL-1F4 Antibody

Catalog # Availability Size / Price Qty
AF2548
AF2548-SP
IL‑18/IL‑1F4  in Human PBMCs.
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Human/Rhesus Macaque IL-18/IL-1F4 Antibody Summary

Species Reactivity
Human, Rhesus Macaque
Specificity
Detects rhesus macaque IL‑18/IL-1F4 in direct ELISAs and Western blots. In Western blots, approximately 10% cross-reactivity with recombinant mouse IL-18, recombinant rat IL-18, and recombinant porcine IL-18 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant rhesus macaque IL‑18/IL-1F4
Tyr37-Asp193
Accession # AAK13416
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Rhesus Macaque IL‑18/IL‑1F4 (Catalog # 2548-RM)
ELISA

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.

 
Immunocytochemistry
5-15 µg/mL
See below
Knockout Validated
IL‑18/IL‑1F4 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in IL‑18/IL‑1F4 knockout HeLa cell line.
 
Neutralization
Measured by its ability to neutralize IL‑18/IL‑1F4-induced IFN‑ gamma secretion in the KG‑1 human acute myelogenous leukemia cell line. Novick, D. et al. (1999) Immunity 10(1):127. The Neutralization Dose (ND50) is typically 0.4‑1.2 µg/mL in the presence of 10 ng/mL Recombinant Rhesus Macaque IL‑18/IL‑1F4 and 20 ng/mL Recombinant Human TNF‑ alpha.

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.

Data Examples

Immunocytochemistry IL‑18/IL‑1F4  in Human PBMCs. View Larger

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.

Neutralization IFN‑ gamma  Secretion Induced by IL‑18/IL‑1F4 and Neutralization by Primate IL‑18/IL‑1F4 Antibody. View Larger

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.

Knockout Validated Western  Blot Shows Human IL‑18/IL‑1F4 Specificity by Using  Knockout Cell Line. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
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.

Background: IL-18/IL-1F4

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).

References
  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.
    Long Name
    Interleukin 18
    Entrez Gene IDs
    3606 (Human); 16173 (Mouse); 29197 (Rat); 397057 (Porcine); 574151 (Primate)
    Alternate Names
    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

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