Anti-Interleukin-18 IL18 Antibody Picoband®

Western blot analysis of IL18 using anti-IL18 antibody (RP1017).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: mouse IL18 recombinant protein,
Lane 2: mouse J774A.1 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-IL18 antigen affinity purified polyclonal antibody (Catalog # RP1017) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for IL18 at approximately 22 kDa.

IHC analysis of IL18 using anti-IL18 antibody (RP1017).
IL18 was detected in a paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-IL18 Antibody (RP1017) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Fgl2 disruption inhibited activation of the NLRP3 inflammasome in NASH. Total protein was obtained from liver tissues of MCD-fed or HFD-fed WT and fgl2-/- mice. MCS-fed and chow-fed mice were used as controls. NLRP3, pro-caspase-1, cleaved caspase-1 (caspase-1 p10), pro-IL-1β, mature IL-1β and IL-18 were analyzed by western blotting (A). BMDMs from WT and fgl2-/- mice were stimulated with LPS or FFA and tested for inflammasomes by western blotting (B). Image density was quantified using ImageLab software. For bar graphs, n=6 in each group. The data represent the mean ± SD from at least three independent experiments. Statistical differences were determined by two-way ANOVA. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; ns, not significant.
Index in PubMed under a CC BY license. PMID: 32863955

Fgl2 deficiency reduced lipid accumulation in hepatocytes by inhibiting the secretion of proinflammatory cytokines in macrophages. BMDMs from WT and fgl2-/- mice were stimulated with LPS (100 ng/ml) or FFA (800 μmol/L). The levels of proinflammatory cytokines, including TNF-α, MCP-1, IL-6, IL-1β and IL-18, in the supernatant of cell cultures were tested by ELISAs (A). Primary hepatocytes were isolated from C57BL/6J livers and incubated with LPS- or FFA-BMDM-CM for 24 hours. The brief experimental procedure is shown in a diagram. Oil red O staining was used to detect fat deposition in primary hepatocytes after treatment with BMDM-CM (B). Then, the mRNA levels of genes involved in lipogenesis (Fasn, SREBP-2) (C) or lipolysis (PPARα, CPT1A) (D) in primary hepatocytes were tested by real-time PCR. For bar graphs, n=6 in each group. The data represent the mean ± SD from at least three independent experiments. Statistical differences were determined by two-way ANOVA. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; ns, not significant.
Index in PubMed under a CC BY license. PMID: 32863955

Fgl2 deficiency attenuated liver inflammatory injury in NASH mice. In MCD-fed or HFD-fed WT and fgl2-/- mice, HE staining was performed to detect histological changes in the liver (A). The NAFLD activity score was evaluated (B). BMDMs were isolated from WT or fgl2-/- mice and injected into macrophage-depleted WT NASH mice (C). Histological changes were detected by HE staining (D, arrows indicate inflammatory infiltration). Serum ALT, AST, LDH (E) and fasting glucose (F) were tested by an automatic biochemical analyzer (n=10 in each group). The levels of serum insulin were tested by an ELISA kit (F). The levels of the proinflammatory cytokines TNF-α, MCP-1, IL-6, IL-1β and IL-18 in the liver were tested by ELISAs (G). For bar graphs, n=6-10 in each group. The data represent the mean ± SD from at least three independent experiments. Statistical differences were determined by two-way ANOVA. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; ns, not significant.
Index in PubMed under a CC BY license. PMID: 32863955