Human/Rhesus Macaque IL‑18/IL‑1F4 Antibody

IFN‑ gamma Secretion Induced by IL‑18/IL‑1F4 and Neutralization by Primate IL‑18/IL‑1F4 Antibody.

In the presence of Recombinant Human TNF-alpha (20 ng/mL, Catalog # 210-TA), Recombinant Rhesus Macaque IL-18/IL-1F4 (Catalog # 2548-RM) stimulates IFN-gamma secretion in the KG-1 human acute myelogenous leukemia cell line in a dose-dependent manner (orange line), as measured by the Human IFN-gamma Quantikine ELISA Kit (Catalog # DIF50C). Under these conditions, IFN-gamma secretion elicited by Recombinant Rhesus Macaque IL-18/IL-1F4 (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human/Rhesus Macaque IL-18/IL-1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548). The ND₅₀ is typically 0.4-1.2 µg/mL.

Western Blot Shows Human IL‑18/IL‑1F4 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and IL-18/IL-1F4 knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Rhesus Macaque IL-18/IL-1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for IL-18/IL-1F4 at approximately 25 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

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

Characteristics of CSC and EMT events regulated by RanBP1 in glioma. (A) Comparison of the expression of GSC marker proteins by RanBP1 gene suppression in GSCs. U87 cells are differentiated into GSCs using TM. (B) Comparison of the marker protein expression according to the expression of RanBP1 using ICC in GSCs. (C) Comparison of differences in sphere formation according to the expression level of RanBP1 in GSCs. Experiments performed in triplicate. (D) Comparison of the EMT marker protein expression after RanBP1 gene suppression using si-RNA. (E) Confirmation of EMT marker protein expression according to the expression of RanBP1 using ICC. (F) Analysis of cell migration and invasion ability after the suppression of RanBP1 expression using si-RNA. (G) Analysis of regulation of IL-18 expression by RanBP1 in GSCs. Error bars represent mean ± SD. Triplicate samples. * p < 0.001, ** p < 0.0005, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Cytokines regulated by RanBP1 in lung cancer. (A) Identification of secreted factors regulated by RanBP1 using cytokine arrays. A549 cells are used, and the expression of RanBP1 is suppressed using si-RNA. (B) Comparison of the gene expression of cytokines regulated by RanBP1. (C) Analysis of cytokines regulated by RanBP1 using WB. (D) Comparison of the sphere formation ability after each neutralizing antibody treatment. The p value is less than 0.0001 versus the IgG Ab value. (E) Comparison of migration and invasion ability of cells after each neutralizing antibody treatment. Experiments performed in triplicate. (F) CSC marker protein analysis in A549 after treatment with IL-18 neutralizing antibody. (G) Confirmation of the expression levels of EMT marker proteins after treatment with the IL-18 neutralizing antibody. (H) Expression analysis of RanBP1 regulated by IL-18 using neutralizing antibodies. Error bars represent mean ± SD. Triplicate samples. * p < 0.05, ** p < 0.001, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Cytokines regulated by RanBP1 in lung cancer. (A) Identification of secreted factors regulated by RanBP1 using cytokine arrays. A549 cells are used, and the expression of RanBP1 is suppressed using si-RNA. (B) Comparison of the gene expression of cytokines regulated by RanBP1. (C) Analysis of cytokines regulated by RanBP1 using WB. (D) Comparison of the sphere formation ability after each neutralizing antibody treatment. The p value is less than 0.0001 versus the IgG Ab value. (E) Comparison of migration and invasion ability of cells after each neutralizing antibody treatment. Experiments performed in triplicate. (F) CSC marker protein analysis in A549 after treatment with IL-18 neutralizing antibody. (G) Confirmation of the expression levels of EMT marker proteins after treatment with the IL-18 neutralizing antibody. (H) Expression analysis of RanBP1 regulated by IL-18 using neutralizing antibodies. Error bars represent mean ± SD. Triplicate samples. * p < 0.05, ** p < 0.001, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Cytokines regulated by RanBP1 in lung cancer. (A) Identification of secreted factors regulated by RanBP1 using cytokine arrays. A549 cells are used, and the expression of RanBP1 is suppressed using si-RNA. (B) Comparison of the gene expression of cytokines regulated by RanBP1. (C) Analysis of cytokines regulated by RanBP1 using WB. (D) Comparison of the sphere formation ability after each neutralizing antibody treatment. The p value is less than 0.0001 versus the IgG Ab value. (E) Comparison of migration and invasion ability of cells after each neutralizing antibody treatment. Experiments performed in triplicate. (F) CSC marker protein analysis in A549 after treatment with IL-18 neutralizing antibody. (G) Confirmation of the expression levels of EMT marker proteins after treatment with the IL-18 neutralizing antibody. (H) Expression analysis of RanBP1 regulated by IL-18 using neutralizing antibodies. Error bars represent mean ± SD. Triplicate samples. * p < 0.05, ** p < 0.001, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Cytokines regulated by RanBP1 in lung cancer. (A) Identification of secreted factors regulated by RanBP1 using cytokine arrays. A549 cells are used, and the expression of RanBP1 is suppressed using si-RNA. (B) Comparison of the gene expression of cytokines regulated by RanBP1. (C) Analysis of cytokines regulated by RanBP1 using WB. (D) Comparison of the sphere formation ability after each neutralizing antibody treatment. The p value is less than 0.0001 versus the IgG Ab value. (E) Comparison of migration and invasion ability of cells after each neutralizing antibody treatment. Experiments performed in triplicate. (F) CSC marker protein analysis in A549 after treatment with IL-18 neutralizing antibody. (G) Confirmation of the expression levels of EMT marker proteins after treatment with the IL-18 neutralizing antibody. (H) Expression analysis of RanBP1 regulated by IL-18 using neutralizing antibodies. Error bars represent mean ± SD. Triplicate samples. * p < 0.05, ** p < 0.001, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Cytokines regulated by RanBP1 in lung cancer. (A) Identification of secreted factors regulated by RanBP1 using cytokine arrays. A549 cells are used, and the expression of RanBP1 is suppressed using si-RNA. (B) Comparison of the gene expression of cytokines regulated by RanBP1. (C) Analysis of cytokines regulated by RanBP1 using WB. (D) Comparison of the sphere formation ability after each neutralizing antibody treatment. The p value is less than 0.0001 versus the IgG Ab value. (E) Comparison of migration and invasion ability of cells after each neutralizing antibody treatment. Experiments performed in triplicate. (F) CSC marker protein analysis in A549 after treatment with IL-18 neutralizing antibody. (G) Confirmation of the expression levels of EMT marker proteins after treatment with the IL-18 neutralizing antibody. (H) Expression analysis of RanBP1 regulated by IL-18 using neutralizing antibodies. Error bars represent mean ± SD. Triplicate samples. * p < 0.05, ** p < 0.001, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Cytokines regulated by RanBP1 in lung cancer. (A) Identification of secreted factors regulated by RanBP1 using cytokine arrays. A549 cells are used, and the expression of RanBP1 is suppressed using si-RNA. (B) Comparison of the gene expression of cytokines regulated by RanBP1. (C) Analysis of cytokines regulated by RanBP1 using WB. (D) Comparison of the sphere formation ability after each neutralizing antibody treatment. The p value is less than 0.0001 versus the IgG Ab value. (E) Comparison of migration and invasion ability of cells after each neutralizing antibody treatment. Experiments performed in triplicate. (F) CSC marker protein analysis in A549 after treatment with IL-18 neutralizing antibody. (G) Confirmation of the expression levels of EMT marker proteins after treatment with the IL-18 neutralizing antibody. (H) Expression analysis of RanBP1 regulated by IL-18 using neutralizing antibodies. Error bars represent mean ± SD. Triplicate samples. * p < 0.05, ** p < 0.001, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Cytokines regulated by RanBP1 in lung cancer. (A) Identification of secreted factors regulated by RanBP1 using cytokine arrays. A549 cells are used, and the expression of RanBP1 is suppressed using si-RNA. (B) Comparison of the gene expression of cytokines regulated by RanBP1. (C) Analysis of cytokines regulated by RanBP1 using WB. (D) Comparison of the sphere formation ability after each neutralizing antibody treatment. The p value is less than 0.0001 versus the IgG Ab value. (E) Comparison of migration and invasion ability of cells after each neutralizing antibody treatment. Experiments performed in triplicate. (F) CSC marker protein analysis in A549 after treatment with IL-18 neutralizing antibody. (G) Confirmation of the expression levels of EMT marker proteins after treatment with the IL-18 neutralizing antibody. (H) Expression analysis of RanBP1 regulated by IL-18 using neutralizing antibodies. Error bars represent mean ± SD. Triplicate samples. * p < 0.05, ** p < 0.001, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Characteristics of CSC and EMT events regulated by RanBP1 in glioma. (A) Comparison of the expression of GSC marker proteins by RanBP1 gene suppression in GSCs. U87 cells are differentiated into GSCs using TM. (B) Comparison of the marker protein expression according to the expression of RanBP1 using ICC in GSCs. (C) Comparison of differences in sphere formation according to the expression level of RanBP1 in GSCs. Experiments performed in triplicate. (D) Comparison of the EMT marker protein expression after RanBP1 gene suppression using si-RNA. (E) Confirmation of EMT marker protein expression according to the expression of RanBP1 using ICC. (F) Analysis of cell migration and invasion ability after the suppression of RanBP1 expression using si-RNA. (G) Analysis of regulation of IL-18 expression by RanBP1 in GSCs. Error bars represent mean ± SD. Triplicate samples. * p < 0.001, ** p < 0.0005, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Cytokines regulated by RanBP1 in lung cancer. (A) Identification of secreted factors regulated by RanBP1 using cytokine arrays. A549 cells are used, and the expression of RanBP1 is suppressed using si-RNA. (B) Comparison of the gene expression of cytokines regulated by RanBP1. (C) Analysis of cytokines regulated by RanBP1 using WB. (D) Comparison of the sphere formation ability after each neutralizing antibody treatment. The p value is less than 0.0001 versus the IgG Ab value. (E) Comparison of migration and invasion ability of cells after each neutralizing antibody treatment. Experiments performed in triplicate. (F) CSC marker protein analysis in A549 after treatment with IL-18 neutralizing antibody. (G) Confirmation of the expression levels of EMT marker proteins after treatment with the IL-18 neutralizing antibody. (H) Expression analysis of RanBP1 regulated by IL-18 using neutralizing antibodies. Error bars represent mean ± SD. Triplicate samples. * p < 0.05, ** p < 0.001, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Cytokines regulated by RanBP1 in lung cancer. (A) Identification of secreted factors regulated by RanBP1 using cytokine arrays. A549 cells are used, and the expression of RanBP1 is suppressed using si-RNA. (B) Comparison of the gene expression of cytokines regulated by RanBP1. (C) Analysis of cytokines regulated by RanBP1 using WB. (D) Comparison of the sphere formation ability after each neutralizing antibody treatment. The p value is less than 0.0001 versus the IgG Ab value. (E) Comparison of migration and invasion ability of cells after each neutralizing antibody treatment. Experiments performed in triplicate. (F) CSC marker protein analysis in A549 after treatment with IL-18 neutralizing antibody. (G) Confirmation of the expression levels of EMT marker proteins after treatment with the IL-18 neutralizing antibody. (H) Expression analysis of RanBP1 regulated by IL-18 using neutralizing antibodies. Error bars represent mean ± SD. Triplicate samples. * p < 0.05, ** p < 0.001, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Cytokines regulated by RanBP1 in lung cancer. (A) Identification of secreted factors regulated by RanBP1 using cytokine arrays. A549 cells are used, and the expression of RanBP1 is suppressed using si-RNA. (B) Comparison of the gene expression of cytokines regulated by RanBP1. (C) Analysis of cytokines regulated by RanBP1 using WB. (D) Comparison of the sphere formation ability after each neutralizing antibody treatment. The p value is less than 0.0001 versus the IgG Ab value. (E) Comparison of migration and invasion ability of cells after each neutralizing antibody treatment. Experiments performed in triplicate. (F) CSC marker protein analysis in A549 after treatment with IL-18 neutralizing antibody. (G) Confirmation of the expression levels of EMT marker proteins after treatment with the IL-18 neutralizing antibody. (H) Expression analysis of RanBP1 regulated by IL-18 using neutralizing antibodies. Error bars represent mean ± SD. Triplicate samples. * p < 0.05, ** p < 0.001, versus control. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047826), licensed under a CC-BY license. Not internally tested by R&D Systems.