Human IL-8/CXCL8 Antibody Best Seller

Chemotaxis Induced by IL-8/CXCL8 and Neutralization by Human IL-8/CXCL8 Antibody.

Recombinant Human IL-8/CXCL8 (Catalog # 208-IL) chemoattracts the BaF3 mouse pro-B cell line transfected with human CXCR2 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Human IL-8/CXCL8 (20 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-8/CXCL8 Monoclonal Antibody (Catalog # MAB208). The ND₅₀ is typically 0.08-0.4 µg/mL.

Detection of IL-8/CXCL8 by Western Blot

CDK1/CDC14B-dependent phosphorylation of USP9X at serine 2563 promotes mitotic survival via WT1 and IL-8. e Immunoblot analysis confirming reversal of mitotic apoptosis following exogenous reconstitution of IL-8 in CXCL8-depleted cells. Experiment was performed as in a, with addition of exogenous IL-8 for the last 48 h. Cells were treated with nocodazole for 8 h. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32152317), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-8/CXCL8 by Western Blot

CDK1/CDC14B-dependent phosphorylation of USP9X at serine 2563 promotes mitotic survival via WT1 and IL-8. f Immunoblot analysis detecting induction of mitotic apoptosis in response to WT1, CXCL8, or WT1 and CXCL8 knockdown compared to control knockdown in U2OS cells that were arrested in mitosis using nocodazole (8 h). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32152317), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-8/CXCL8 by Western Blot

CDK1/CDC14B-dependent phosphorylation of USP9X at serine 2563 promotes mitotic survival via WT1 and IL-8.a. i Immunoblot analysis revealing decreased mitotic apoptosis after CDC14B knockdown in USP9XWT but not in USP9XMut U2OS cells. Before lysis cells were transfected with control or CDC14B-directed siRNA, arrested in mitosis, and collected for analysis by western blot. Throughout this figure, mean and standard deviations as error bars are displayed. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32152317), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-8/CXCL8 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 IL-8/CXCL8 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 IL-8/CXCL8 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 IL-8/CXCL8 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 IL-8/CXCL8 by Flow Cytometry

Interaction between osteosarcoma and macrophages increased IL-8 production. A Cytokine arrays of 143B-Luc cells, human macrophages, and co-culture CM. B IL-8 production during the co-culture of OS cells (143B-Luc, SJSA-1) and macrophages (THP-1 Mφ, HMDMs). C IL-8 expression in macrophages stimulated with OS-CM. D IL-8 expression in OS cells stimulated with macrophage CM. E UMAP plot of OS lung metastases. F UMAP plot of Iba1, CD163, and IL-8 expression in myeloid cell clusters. G Violin plot depicting IL-8 expression in Iba1+/− or CD163+/− myeloid cell clusters. CM conditioned medium, HMDMs human monocyte-derived macrophages, OS osteosarcoma, THP-1 Mφ THP-1-derived macrophage. Data are presented as mean ± standard deviation. **p < 0.01. All data were obtained from at least three independent experiments. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41419-024-06487-y), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-8/CXCL8 by Immunohistochemistry

Effect of IL-8 in co-culture CM on proliferation, migration, and invasion of osteosarcoma cells.A Cell viability assay for OS cells treated with or without recombinant IL-8 (rIL-8, 10 ng/mL) using Cell Counting Kit-8. B, C Scratch and invasion assays for OS cells treated with or without co-culture CM. The scratch assay was performed for 12 h, and the invasion assay was conducted for 24 h. Scale bars represent 500 μm. D Cell viability assay for OS cells treated with or without co-culture CM and with or without anti-IL-8 antibodies (IL-8 Abs, 1 µg/mL) using Cell Counting Kit-8. E, F Scratch and invasion assays for OS cells treated with or without co-culture CM and with or without IL-8 Abs (100 ng/mL). The scratch assay was performed for 12 h, and the invasion assay was conducted for 24 h. Scale bars represent 500 μm. CM: conditioned medium, OS: osteosarcoma, ns: not significant. Data are presented as mean ± standard deviation. *p < 0.05 and **p < 0.01. All data were obtained from at least three independent experiments. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41419-024-06487-y), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-8/CXCL8 by Immunohistochemistry

Effect of IL-8 in co-culture CM on lung metastasis. A Schematic showing the experimental design for OS tail vein injection. 143B-Luc cells were treated with DMEM or co-culture CM for 12 h prior to tail vein injection; 143B-Luc cells (1 × 106/mouse) were injected into the tail vein of 6–7 week-old BALB/c nu/nu mice. B IVIS imaging and quantification of lung metastasis 14 days after the injection of tumors pretreated with or without co-culture CM. C Hematoxylin and eosin staining of lung sections with or without co-culture CM and quantification of lung colonies. Scale bars represent 1 mm and 500 µm. D Immunohistochemistry of Ki-67 positive cells in lung colonies with or without co-culture CM and quantification of the Ki-67 labeling index. Two colonies per sample were randomly selected, and quantification was performed for 10 colonies. Ki-67-positive cells in the colonies were counted and shown as the Ki-67 labeling index. Scale bars represent 100 µm. E Schematic showing the experimental design for OS tail vein injection using anti-IL-8 antibody. F IVIS imaging and quantification of lung metastasis 14 days after the injection of tumors pretreated with co-culture CM with or without IL-8 antibodies. G Hematoxylin and eosin staining of lung sections with co-culture CM with or without IL-8 Abs and quantification of lung colonies. Scale bars represent 1 mm and 500 µm. H Immunohistochemistry of Ki-67-positive cells in lung colonies with co-culture CM with or without IL-8 Abs and quantification of Ki-67 labeling index. Scale bars represent 100 µm. CM conditioned medium, DMEM Dulbecco’s modified Eagle’s medium, IVIS, in vivo imaging system, OS osteosarcoma. Data are presented as mean ± standard deviation; *p < 0.05, **p < 0.01. Each group contained five animals. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41419-024-06487-y), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-8/CXCL8 by Immunohistochemistry

Effect of IL-8 in co-culture CM on primary growth. A Schematic showing the experimental design for subcutaneous transplantation of OS cells. 143B-Luc cells (2 × 106/mouse) were subcutaneously transplanted in 6–7-week-old BALB/c nu/nu mice, and DMEM or co-culture CM was injected into the para-tumor thrice per week. B Tumor volume with or without co-culture CM measured thrice per week after tumor transplantation. C, D Tumor weight and image of the excised tumor with or without co-cultured CM 14 days after tumor transplantation. E, F Immunohistochemistry and quantification of Ki-67 and phospho-FAK in tumor sections with or without co-culture CM 14 days after tumor transplantation. Two fields of view per sample were randomly selected, and quantification was performed in 10 fields. E Ki-67-positive cells in the field were counted and indicated as Ki-67 labeling index. Scale bars represent 50 μm. G Schematic showing the experimental design for subcutaneous OS transplantation using anti-IL-8 antibodies (IL-8 Abs). H Tumor volume after pretreatment with co-culture CM with or without IL-8 Abs (10 µg/mouse) measured thrice per week after tumor transplantation. I and J Weight and image of the excised tumor pretreated with co-culture CM with or without IL-8 Abs 14 days after tumor transplantation. K, L Immunohistochemistry and quantification of Ki-67 and phospho-FAK in tumor sections with co-culture CM with or without IL-8 Abs 14 days after tumor transplantation. Scale bars represent 50 µm. CM conditioned medium, DMEM Dulbecco’s modified Eagle’s medium, FAK focal adhesion kinase, OS osteosarcoma. Data are presented as mean ± standard deviation; *p < 0.05, **p < 0.01. Each group contained five animals. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41419-024-06487-y), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-8/CXCL8 by Flow Cytometry

Interaction between osteosarcoma and macrophages increased IL-8 production. A Cytokine arrays of 143B-Luc cells, human macrophages, and co-culture CM. B IL-8 production during the co-culture of OS cells (143B-Luc, SJSA-1) and macrophages (THP-1 Mφ, HMDMs). C IL-8 expression in macrophages stimulated with OS-CM. D IL-8 expression in OS cells stimulated with macrophage CM. E UMAP plot of OS lung metastases. F UMAP plot of Iba1, CD163, and IL-8 expression in myeloid cell clusters. G Violin plot depicting IL-8 expression in Iba1+/− or CD163+/− myeloid cell clusters. CM conditioned medium, HMDMs human monocyte-derived macrophages, OS osteosarcoma, THP-1 Mφ THP-1-derived macrophage. Data are presented as mean ± standard deviation. **p < 0.01. All data were obtained from at least three independent experiments. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41419-024-06487-y), licensed under a CC-BY license. Not internally tested by R&D Systems.

Human IL-8 / CXCL8 ELISA Standard Curve

Recombinant Human IL-8/CXCL8 (Catalog # 208-IL) was serially diluted and captured by Mouse Anti-Human IL-8/CXCL8 Monoclonal Antibody (Catalog # MAB208) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human IL-8/CXCL8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-208-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)