Key Product Details

Species Reactivity

Validated:

Rat

Cited:

Rat

Applications

Validated:

Western Blot, Neutralization

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Binding Assay, ELISA Development

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all IL-18/IL-1F4 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant rat IL‑18/IL-1F4
His37-Ser194
Accession # P97636

Specificity

Detects rat IL‑18/IL-1F4 in direct ELISAs and Western blots. In Western blots, approximately 35% cross-reactivity with recombinant mouse IL‑18 is observed and approximately 10% cross-reactivity with recombinant human 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 Rat IL‑18/IL‑1F4 Antibody

IFN‑ gamma Secretion Induced by IL‑18/IL‑1F4 and Neutralization by Rat IL‑18/IL‑1F4 Antibody.

IFN‑ gamma Secretion Induced by IL‑18/IL‑1F4 and Neutralization by Rat IL‑18/IL‑1F4 Antibody.

In the presence of Recombinant Mouse IL-12 (0.1 ng/mL, Catalog # 419-ML), Recombinant Rat IL-18/IL-1F4 (Catalog # 521-RL) stimulates IFN-gamma secretion in activated mouse T cells in a dose-dependent manner (orange line), as measured by the Mouse IFN-gamma Quantikine ELISA Kit (Catalog # MIF00). Under these conditions, IFN-gamma secretion elicited by Recombinant Rat IL-18/IL-1F4 (15 ng/mL) is neutralized (green line) by increasing concentrations of Rat IL-18/IL-1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF521). The ND50 is typically 1-3 µg/mL.
Detection of IL-18/IL-1F4 by Western Blot

Detection of IL-18/IL-1F4 by Western Blot

Effects of pirfenidone (PFD) on NLRP3 inflammasome activation in PAH lungs and in macrophages in vitro: Lung tissue was isolated from the rats as described in Figures 1, 2, 3, homogenized and cleavage of caspase‐1 (a, b), IL‐1 beta (c, d), and IL‐18 (e, f) were assessed by Western blot. Representative pictures are shown (a, c, e), and quantified (b, d, f). (g, h) Bone marrow was isolated from mice and differentiated into bone marrow‐derived macrophages (BMDMs) in DMEM supplemented with a 20% l‐cell conditioned medium for 7 days. Subsequently, BMDMs were treated with or without PFD (500 µg/ml) o/n, and subsequently with lipopolysaccharide (LPS) (100 ng/ml) for 4 h. (g) Cells were lysed, RNA was isolated, and the expression of NLRP3, pro‐IL‐1 beta, and pro‐IL‐18 mRNA was measured by qPCR and corrected for 36B4, cyclophilin B, and GAPDH (housekeeping genes). (h) Cells were subsequently treated with nigericin (NG) (20 µM) for 30 min and IL‐1 beta secretion into the medium was assessed by ELISA and corrected for cell protein; *p < 0.05; **p < 0.01; ***p < 0.001 by two‐sided one‐way ANOVA with Holm–Sidak's post hoc correction. ANOVA, analysis of variance; ELISA, enzyme‐linked immunosorbent assay; IL‐1 beta, interleukin‐1 beta ; mRNA, messenger RNA; PAH, pulmonary arterial hypertension; qPCR, quantitative polymerase chain reaction. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35833096), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of IL-18/IL-1F4 by Western Blot

Detection of IL-18/IL-1F4 by Western Blot

Effects of pirfenidone (PFD) on NLRP3 inflammasome activation in PAH lungs and in macrophages in vitro: Lung tissue was isolated from the rats as described in Figures 1, 2, 3, homogenized and cleavage of caspase‐1 (a, b), IL‐1 beta (c, d), and IL‐18 (e, f) were assessed by Western blot. Representative pictures are shown (a, c, e), and quantified (b, d, f). (g, h) Bone marrow was isolated from mice and differentiated into bone marrow‐derived macrophages (BMDMs) in DMEM supplemented with a 20% l‐cell conditioned medium for 7 days. Subsequently, BMDMs were treated with or without PFD (500 µg/ml) o/n, and subsequently with lipopolysaccharide (LPS) (100 ng/ml) for 4 h. (g) Cells were lysed, RNA was isolated, and the expression of NLRP3, pro‐IL‐1 beta, and pro‐IL‐18 mRNA was measured by qPCR and corrected for 36B4, cyclophilin B, and GAPDH (housekeeping genes). (h) Cells were subsequently treated with nigericin (NG) (20 µM) for 30 min and IL‐1 beta secretion into the medium was assessed by ELISA and corrected for cell protein; *p < 0.05; **p < 0.01; ***p < 0.001 by two‐sided one‐way ANOVA with Holm–Sidak's post hoc correction. ANOVA, analysis of variance; ELISA, enzyme‐linked immunosorbent assay; IL‐1 beta, interleukin‐1 beta ; mRNA, messenger RNA; PAH, pulmonary arterial hypertension; qPCR, quantitative polymerase chain reaction. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35833096), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of IL-18/IL-1F4 by Western Blot

Detection of IL-18/IL-1F4 by Western Blot

Effects of pirfenidone (PFD) on NLRP3 inflammasome activation in PAH lungs and in macrophages in vitro: Lung tissue was isolated from the rats as described in Figures 1, 2, 3, homogenized and cleavage of caspase‐1 (a, b), IL‐1 beta (c, d), and IL‐18 (e, f) were assessed by Western blot. Representative pictures are shown (a, c, e), and quantified (b, d, f). (g, h) Bone marrow was isolated from mice and differentiated into bone marrow‐derived macrophages (BMDMs) in DMEM supplemented with a 20% l‐cell conditioned medium for 7 days. Subsequently, BMDMs were treated with or without PFD (500 µg/ml) o/n, and subsequently with lipopolysaccharide (LPS) (100 ng/ml) for 4 h. (g) Cells were lysed, RNA was isolated, and the expression of NLRP3, pro‐IL‐1 beta, and pro‐IL‐18 mRNA was measured by qPCR and corrected for 36B4, cyclophilin B, and GAPDH (housekeeping genes). (h) Cells were subsequently treated with nigericin (NG) (20 µM) for 30 min and IL‐1 beta secretion into the medium was assessed by ELISA and corrected for cell protein; *p < 0.05; **p < 0.01; ***p < 0.001 by two‐sided one‐way ANOVA with Holm–Sidak's post hoc correction. ANOVA, analysis of variance; ELISA, enzyme‐linked immunosorbent assay; IL‐1 beta, interleukin‐1 beta ; mRNA, messenger RNA; PAH, pulmonary arterial hypertension; qPCR, quantitative polymerase chain reaction. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35833096), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of IL-18/IL-1F4 by Western Blot

Detection of IL-18/IL-1F4 by Western Blot

Effects of pirfenidone (PFD) on NLRP3 inflammasome activation in PAH lungs and in macrophages in vitro: Lung tissue was isolated from the rats as described in Figures 1, 2, 3, homogenized and cleavage of caspase‐1 (a, b), IL‐1 beta (c, d), and IL‐18 (e, f) were assessed by Western blot. Representative pictures are shown (a, c, e), and quantified (b, d, f). (g, h) Bone marrow was isolated from mice and differentiated into bone marrow‐derived macrophages (BMDMs) in DMEM supplemented with a 20% l‐cell conditioned medium for 7 days. Subsequently, BMDMs were treated with or without PFD (500 µg/ml) o/n, and subsequently with lipopolysaccharide (LPS) (100 ng/ml) for 4 h. (g) Cells were lysed, RNA was isolated, and the expression of NLRP3, pro‐IL‐1 beta, and pro‐IL‐18 mRNA was measured by qPCR and corrected for 36B4, cyclophilin B, and GAPDH (housekeeping genes). (h) Cells were subsequently treated with nigericin (NG) (20 µM) for 30 min and IL‐1 beta secretion into the medium was assessed by ELISA and corrected for cell protein; *p < 0.05; **p < 0.01; ***p < 0.001 by two‐sided one‐way ANOVA with Holm–Sidak's post hoc correction. ANOVA, analysis of variance; ELISA, enzyme‐linked immunosorbent assay; IL‐1 beta, interleukin‐1 beta ; mRNA, messenger RNA; PAH, pulmonary arterial hypertension; qPCR, quantitative polymerase chain reaction. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35833096), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Rat IL‑18/IL‑1F4 Antibody

Application
Recommended Usage

Western Blot

0.1 µg/mL
Sample: Recombinant Rat IL‑18/IL‑1F4 (Catalog # 521-RL)

Neutralization

Measured by its ability to neutralize IL‑18/IL‑1F4-induced IFN‑ gamma secretion in activated mouse T cells [Ahn, H.J. et al. (1997) J. Immunol. 159:2125]. The Neutralization Dose (ND50) is typically 1-3 µg/mL in the presence of 15 ng/mL Recombinant Rat IL‑18/IL‑1F4 and 0.1 ng/mL Recombinant Mouse IL‑12.

Reviewed Applications

Read 1 review rated 4 using AF521 in the following applications:

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.

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.

Calculators

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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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). Rat IL-18 is synthesized as a 24 kDa proprotein that contains a 36 amino acid (aa) propeptide and a 158 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 rat IL-18 shares 91% aa sequence identity with mouse IL-18 and 60-64% aa sequence identity with human, canine, feline, porcine, and rhesus macaque 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. Culhane, A.C. et al. (1998) Mol. Psychiatry 3:362.
  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

P97636

Additional IL-18/IL-1F4 Products

Product Documents for Rat IL‑18/IL‑1F4 Antibody

Certificate of Analysis

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Product Specific Notices for Rat IL‑18/IL‑1F4 Antibody

For research use only

Citations for Rat IL‑18/IL‑1F4 Antibody

Customer Reviews for Rat IL‑18/IL‑1F4 Antibody (1)

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  • Rat IL‑18/IL‑1F4 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Rat,spinal cord dorsal horn, particle nerve injury (CCI)
    Verified Customer | Posted 10/26/2015
    Fixed with 4%Polyoxymethylene and 15%saturation picric acid. Without antigen retrieval. <br/> Frozen section, 20m, floating slice method.<br/> First antibody 4 degrees over night and secondary antibody 2 hours at room-temperature.<br/> Antigen retrieval in 99 degrees citrate buffer(pH 4.6) for 5min would benefit the output (not tested).<br/> Green: Interleukin-18; red: Iba-1 (microglia marker). <br />Specificity: Reasonably specific<br />Sensitivity: Reasonably sensitive<br />Buffer: PBS<br />Dilution: 1/100

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