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

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

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.