Key Product Details

Species Reactivity

Validated:

Porcine

Cited:

Mouse

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Western Blot

Label

Unconjugated

Antibody Source

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

Product Specifications

Immunogen

E. coli-derived recombinant porcine IL‑18/IL‑1F4

Specificity

Detects porcine IL‑18/IL‑1F4 in direct ELISAs and Western blots. In direct ELISAs, approximately 30% cross‑reactivity with recombinant human IL-18 is observed and 10% cross-reactivity with recombinant rat IL‑18 and recombinant mouse IL‑18 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Porcine IL‑18/IL‑1F4 Antibody

IL-18/IL-1F4 antibody in Porcine PBMCs by Immunocytochemistry (ICC).

IL‑18/IL‑1F4 in Porcine PBMCs.

IL-18/IL-1F4 was detected in immersion fixed porcine peripheral blood mononuclear cells (PBMCs) using Goat Anti-Porcine IL-18/IL-1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF588) at 15 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 (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Porcine IL-18/IL-1F4 by Western Blot

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

MA attenuates LPS-induced inflammatory effects through inhibition of NLRP3. (a) qRT-PCR was performed to evaluate the mRNA expression of NLRP3, ASC and Caspase-1 in RAW264.7 cells. (b) Western blot analysis was performed to detect the protein levels of NLRP3, ASC, and Caspase-1 in RAW264.7 cells. (c) qRT-PCR was performed to evaluate the mRNA expression of NLRP3 in RAW264.7 cells after siRNA treatment. Cells were first transfected with siRNA for 48 h, followed by treatment with 100mg/L MA for 12h, and finally treated with 1mg/L LPS for 24 h. All subsequent experimental cell treatments in this figure are performed in the same way as this cell treatment. (d) Western blot analysis was performed to detect the protein levels of NLRP3 in RAW264.7 cells after siRNA treatment. (e) qRT-PCR was performed to evaluate the mRNA expression of IL-1 beta and IL-18 in RAW264.7 cells after siRNA treatment. (f) Western blot was performed to detect the protein levels of IL-1 beta and IL-18 in RAW264.7 cells after siRNA treatment. (g) ELISA was conducted to detect the levels of IL-1 beta inflammatory cytokines after siRNA treatment. (h) qRT-PCR was performed to evaluate the mRNA expression of NLRP3, ASC, and Caspase-1 in RAW264.7 cells after siRNA treatment. (i) Western blot was performed to detect the protein levels of NLRP3, ASC, and Caspase-1 in RAW264.7 cells after siRNA treatment. Data are shown as mean ± SEM obtained from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36293333), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Porcine IL-18/IL-1F4 by Western Blot

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

MA exhibits anti-inflammatory effects in LPS-induced RAW264.7 cells. (a) Cell viability was assessed by CCK-8 assay in RAW264.7 cells treated with different concentrations of MA (0-500 mg/L) for 12 h, followed by 1 mg/L LPS stimulation for 24 h. (b) qRT-PCR was performed to evaluate the mRNA expressions of TNF-alpha, IL-1 beta, IL-6, and IL-18 in RAW264.7 cells after treatment with 100 mg/L MA for 12 h, followed by stimulation with 1 mg/L LPS for 24 h. (c) Western blot was performed to detect the protein levels of IL-1 beta and IL-18 in RAW264.7 cells. (d) ELISA was conducted to detect the levels of inflammatory cytokines including TNF-alpha, IL-1 beta, IL-6, and IL-18 in RAW264.7 cells. Data are shown as mean ± SEM obtained from three independent experiments. ** p < 0.01, *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36293333), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Porcine IL-18/IL-1F4 by Western Blot

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

MA exhibits anti-inflammatory effects in LPS-induced RAW264.7 cells. (a) Cell viability was assessed by CCK-8 assay in RAW264.7 cells treated with different concentrations of MA (0-500 mg/L) for 12 h, followed by 1 mg/L LPS stimulation for 24 h. (b) qRT-PCR was performed to evaluate the mRNA expressions of TNF-alpha, IL-1 beta, IL-6, and IL-18 in RAW264.7 cells after treatment with 100 mg/L MA for 12 h, followed by stimulation with 1 mg/L LPS for 24 h. (c) Western blot was performed to detect the protein levels of IL-1 beta and IL-18 in RAW264.7 cells. (d) ELISA was conducted to detect the levels of inflammatory cytokines including TNF-alpha, IL-1 beta, IL-6, and IL-18 in RAW264.7 cells. Data are shown as mean ± SEM obtained from three independent experiments. ** p < 0.01, *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36293333), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Porcine IL-18/IL-1F4 by Western Blot

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

MA attenuates LPS-induced inflammatory effects through inhibition of NLRP3. (a) qRT-PCR was performed to evaluate the mRNA expression of NLRP3, ASC and Caspase-1 in RAW264.7 cells. (b) Western blot analysis was performed to detect the protein levels of NLRP3, ASC, and Caspase-1 in RAW264.7 cells. (c) qRT-PCR was performed to evaluate the mRNA expression of NLRP3 in RAW264.7 cells after siRNA treatment. Cells were first transfected with siRNA for 48 h, followed by treatment with 100mg/L MA for 12h, and finally treated with 1mg/L LPS for 24 h. All subsequent experimental cell treatments in this figure are performed in the same way as this cell treatment. (d) Western blot analysis was performed to detect the protein levels of NLRP3 in RAW264.7 cells after siRNA treatment. (e) qRT-PCR was performed to evaluate the mRNA expression of IL-1 beta and IL-18 in RAW264.7 cells after siRNA treatment. (f) Western blot was performed to detect the protein levels of IL-1 beta and IL-18 in RAW264.7 cells after siRNA treatment. (g) ELISA was conducted to detect the levels of IL-1 beta inflammatory cytokines after siRNA treatment. (h) qRT-PCR was performed to evaluate the mRNA expression of NLRP3, ASC, and Caspase-1 in RAW264.7 cells after siRNA treatment. (i) Western blot was performed to detect the protein levels of NLRP3, ASC, and Caspase-1 in RAW264.7 cells after siRNA treatment. Data are shown as mean ± SEM obtained from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36293333), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Porcine IL-18/IL-1F4 by Western Blot

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

MAs demonstrate anti-inflammatory properties in H2O2-induced IPEC-1 cells. (A) The viability of IPEC-1 cells was determined using the CCK-8 assay. (B) The gene expression levels of TNF-alpha, IL-1 beta, IL-6, and IL-18 in IPEC-1 cells after treatment with 100 mg/L of MAs for 12 h, followed by stimulation with 1 mM H2O2 for 1 h. (C) The protein expression levels of IL-1 beta and IL-18. (D) The protein levels of TNF-alpha, IL-1 beta, IL-6, and IL-18. * p < 0.05, ** p < 0.01, *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/2076-3921/13/5/533), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed porcine peripheral blood mononuclear cells

Western Blot

0.1 µg/mL
Sample: Recombinant Porcine IL‑18/IL‑1F4 (Catalog # 588-PL)

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). Porcine IL-18 is synthesized as a 24 kDa proprotein that contains a 35 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 porcine IL-18 shares 88% aa sequence identity with canine and feline IL-18 and 61%‑76% with human, mouse, rat,  and rhesus 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, 11, 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. Muneta, Y. et al. (2000) Cytokine 12:566.
  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. 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

Additional IL-18/IL-1F4 Products

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

Certificate of Analysis

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

For research use only

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

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