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

R&D Systems, part of Bio-Techne | Catalog # AF2548

R&D Systems, part of Bio-Techne
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AF2548
AF2548-SP
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Rhesus Macaque

Cited:

Human

Applications

Validated:

ELISA, Immunocytochemistry, Knockout Validated, Neutralization, Simple Western, Western Blot

Cited:

Bioassay, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all IL-18/IL-1F4 Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant rhesus macaque IL‑18/IL-1F4
Tyr37-Asp193
Accession # AAK13416

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human/Rhesus Macaque IL-18/IL-1F4 Antibody

Detection of Human IL-18/IL-1F4 antibody by Western Blot.

Detection of Human IL‑18/IL‑1F4 by Western Blot.

Western blot shows lysates of PC-3 human prostate cancer cell line, MCF 10A human breast epithelial cell line, A431 human epithelial carcinoma cell line, and HEK293T human embryonic kidney cell line (negative control cell line). PVDF membrane was probed with 0.5 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 22 kDa (as indicated). GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
IL-18/IL-1F4 antibody in Human PBMCs by Immunocytochemistry (ICC).

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.
Detection of Human IL-18/IL-1F4 antibody by Simple WesternTM.

Detection of Human IL‑18/IL‑1F4 by Simple WesternTM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma parental cell line and IL-18/IL-1F4 knockout HeLa cell line, loaded at 0.2 mg/mL. A specific band was detected for IL-18/IL-1F4 at approximately 29 kDa (as indicated) in the HeLa parental cell line using 50 µg/mL of Goat Anti-Human/Rhesus Macaque IL-18/IL-1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 2-40. kDa separation system.
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Applications for Human/Rhesus Macaque IL-18/IL-1F4 Antibody

Application
Recommended Usage

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
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs)

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.

Simple Western

50 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line

Western Blot

0.5 µg/mL
Sample: PC‑3 human prostate cancer cell line, MCF 10A human breast epithelial cell line, and A431 human epithelial carcinoma 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.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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

    Alternate Names

    IGIF, IL-1F4, IL-1g, IL18

    Entrez Gene IDs

    3606 (Human); 16173 (Mouse); 29197 (Rat); 397057 (Porcine); 574151 (Primate)

    Gene Symbol

    IL18

    UniProt

    AAK13416

    Additional IL-18/IL-1F4 Products

    Product Documents for Human/Rhesus Macaque IL-18/IL-1F4 Antibody

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