Porcine IL-18/IL-1F4 Antibody

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

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

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.