IL-18 BPc Antibody [Unconjugated]

LPS-induced IL-18 levels are regulated by LXR activation.(a) BMDM were treated with vehicle (DMSO), GW3965 (1 μ mol/L) or LXR antagonist (LXR ant, 1 μ mol/L) for 24 hours with or without LPS (100 ng/ml) for the last 6 hours. IL-18 mRNA levels were analyzed by RT-qPCR. Values indicate expression normalized to cyclophilin and are presented relative to the expression in LPS-treated cells, set as 1. Values represent the mean ± SEM (3–4 different BMDM preparations). t-test: ***p ≤ 0.001, ns p > 0.05. (b) IL-18 mRNA expression in BMDM from WT and LXR alpha beta −/− mice were analyzed by RT-qPCR. Shown is a representative experiment performed in triplicate (mean ± SD) t-test: *p ≤ 0.05, ns p > 0.05. (c) BMDM were treated as in A and then activated with ATP for the last 2 hours. Pro-IL-18 expression was analyzed by immunoblotting and Hsp90 levels were assayed as loading control. Shown is a representative experiment of three. (d) BMDM were treated as in A. Intracellular expression of IL-18 is shown as analyzed by immunoblotting. Samples shown were analysed on non consecutive lanes of the same blot. (e) BMDM were treated as indicated. Secreted IL-18 levels in culture supernatants were quantified by ELISA. Values represent mean ± SEM (n = 3). t-test: *p ≤ 0.05. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/srep25481), licensed under a CC-BY license. Not internally tested by R&D Systems.

Identification of IL-18BP as a novel LXR target gene.(a) BMDM were treated with vehicle (DMSO) or GW3965 (GW) for 24 hours and IL-18BP mRNA content was analyzed by RT-qPCR. Values indicate expression normalized to cyclophilin and are presented relative to the expression in vehicle-treated cells. Values represent the mean of 4 independent experiments ± SEM. t-test: ***p ≤ 0.001. (b) BMDM from WT or LXR alpha beta deficient mice (LXR alpha beta −/−) were treated as in A with or without the RXR activator LG268 (LG). IL-18BP mRNA content was analyzed as in A. Values represent the mean of 3 independent experiments ± SEM. *p ≤ 0.05, **p ≤ 0.01. (c) BMDM were treated with vehicle (DMSO) or GW3965 (GW) for 24 hours supplemented with Brefeldin A for the last 6 hours. Protein content was analyzed by immunoblotting and Hsp90 levels were assayed as loading control. (d) RAW264.7 cells were transfected with hLXR alpha or pcDNA3.1 (− ) along with the indicated IL-18BP luciferase reporters or pGL3 empty vector. Cells were treated as described in the Methods section. For each reporter, luciferase and beta -galactosidase activities were measured and the ratio was compared to the vehicle-treated condition in the absence of LXR alpha (− ), which was set as 1. Data are mean value ± SD (n = 3) of one representative experiment. t-test: **p ≤ 0.01, ***p ≤ 0.001. (e) Upper panel, location of a putative LXRE in the IL-18BP locus. Arrows indicate the position of primers used for ChIP assays. Lower panel, BMDM cells were incubated with or without GW3965 at 1 μ mol/L for 2 hours. LXR occupancy was assessed by ChIP assays. Primers shown in upper panel were used to amplify an unrelated site at − 4.7 kb that served as a negative control, the − 1.1 kb putative LXRE, − 0.5 kb and TSS as GW3965 responsive sites. SREBP1c primers were used as a positive control for LXR binding. Shown is a representative experiment of three. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/srep25481), licensed under a CC-BY license. Not internally tested by R&D Systems.

Identification of IL-18BP as a novel LXR target gene.(a) BMDM were treated with vehicle (DMSO) or GW3965 (GW) for 24 hours and IL-18BP mRNA content was analyzed by RT-qPCR. Values indicate expression normalized to cyclophilin and are presented relative to the expression in vehicle-treated cells. Values represent the mean of 4 independent experiments ± SEM. t-test: ***p ≤ 0.001. (b) BMDM from WT or LXR alpha beta deficient mice (LXR alpha beta −/−) were treated as in A with or without the RXR activator LG268 (LG). IL-18BP mRNA content was analyzed as in A. Values represent the mean of 3 independent experiments ± SEM. *p ≤ 0.05, **p ≤ 0.01. (c) BMDM were treated with vehicle (DMSO) or GW3965 (GW) for 24 hours supplemented with Brefeldin A for the last 6 hours. Protein content was analyzed by immunoblotting and Hsp90 levels were assayed as loading control. (d) RAW264.7 cells were transfected with hLXR alpha or pcDNA3.1 (− ) along with the indicated IL-18BP luciferase reporters or pGL3 empty vector. Cells were treated as described in the Methods section. For each reporter, luciferase and beta -galactosidase activities were measured and the ratio was compared to the vehicle-treated condition in the absence of LXR alpha (− ), which was set as 1. Data are mean value ± SD (n = 3) of one representative experiment. t-test: **p ≤ 0.01, ***p ≤ 0.001. (e) Upper panel, location of a putative LXRE in the IL-18BP locus. Arrows indicate the position of primers used for ChIP assays. Lower panel, BMDM cells were incubated with or without GW3965 at 1 μ mol/L for 2 hours. LXR occupancy was assessed by ChIP assays. Primers shown in upper panel were used to amplify an unrelated site at − 4.7 kb that served as a negative control, the − 1.1 kb putative LXRE, − 0.5 kb and TSS as GW3965 responsive sites. SREBP1c primers were used as a positive control for LXR binding. Shown is a representative experiment of three. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/srep25481), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu
• Applications: WB
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

IL-18 BPc Antibody [Unconjugated] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant mouse IL‑18 BPc
  Arg25-Ser192
  Accession # AAD17193
• Specificity: Detects mouse IL-18 BPc in direct ELISAs and Western blots. In direct ELISAs, approximately 35% cross‑reactivity with recombinant mouse IL-18 BPd is observed and less than 1% cross-reactivity with recombinant human IL-18 BPa is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: Il18r1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for IL-18 BPc Antibody [Unconjugated]

• I;Il1;Il18r1;Il18ralpha;Il1rrp;Interleukin-18 receptor 1
• IL18 BPc
• IL-18 BPc

Background

Interleukin 18 binding protein (IL-18 BP) is a secreted glycoprotein, which functions as an IL-18 antagonist by binding to IL-18 and blocking its biological activity. IL‑18 BP bears no amino acid sequence homology to the membrane-associated IL-18 and IL-1 receptor proteins. The gene for human IL-18 BP has been localized to chromosome 11q13. It encodes for at least four isoforms by alternative splicing. The IL-18 BP isoforms a and c each contain one immunoglobulin (Ig)-like C2-type domain while isoforms b and d lack a complete Ig domain. The complete Ig domain has been shown to be essential to the binding and neutralizing properties of the binding proteins. Two isoforms of mouse IL18 BP (c and d) containing the complete Ig domain have also been isolated and shown to neutralize IL-18 bioactivity. Human and mouse IL-18 BPs share approximately 61% amino acid sequence identity. Several poxviruses also encode proteins with sequence similarity to the human and mouse IL-18 BP. Viral IL-18 BPs have been shown to bind and inhibit IL-18 responses and may be involved in modulating host immune responses. The expression of IL-18 BP is markedly up-regulated by IFN-gamma , suggesting that IL-18 activity is modulated by a negative feedback mechanism mediated by IL-18 BP.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for IL-18 BPc Antibody (AF129)(1)

Publication using AF129

• The nuclear receptor LXR modulates interleukin-18 levels in macrophages through multiple mechanisms Sci Rep, 2016-05-06;6(0):25481. 2016-05-06 [PMID: 27149934] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: Il18r1
• Uniprot:
  • AAD17193()