Human IL-18 R alpha /IL-1 R5 Antibody

IFN‑ gamma Secretion Induced by IL‑18/IL‑1F4 and Neutralization by IL‑18 R alpha /IL‑1 R5 Antibody. In the presence of Recombinant Human TNF-alpha (20 ng/mL, Catalog # 210-TA), Recombinant Human IL-18/IL-1F4 (Catalog # B003-5) stimulates IFN-gamma secretion in KG-1 human myeloid leukemia cells 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 Human IL-18/IL-1F4 (40 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-18 Ra/ IL-1 R5 Monoclonal Antibody (Catalog # MAB840). The ND₅₀ is typically 0.03-0.1 µg/mL.

IL‑18 R alpha /IL‑1 R5 in Human PBMCs. IL-18 Ra/IL-1 R5 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using 10 µg/mL Mouse Anti-Human IL-18 Ra/IL-1 R5 Mono-clonal Antibody (Catalog # MAB840) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of IL‑18 R alpha /IL‑1 R5 in Human Skin. IL‑18 R alpha /IL‑1 R5 was detected in immersion fixed paraffin-embedded sections of Human Skin using Mouse Anti-Human IL‑18 R alpha /IL‑1 R5 Monoclonal Antibody (Catalog # MAB840) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and cell surface of keratinocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human IL-18R alpha/IL-1 R5 by Western Blot Soluble interleukin-18 receptor alpha complex is associated with interleukin-18 and the soluble form of the interleukin-18 receptor beta chain. (A) Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of the serum interleukin (IL)-18 receptor alpha (IL-18R alpha ) complex. Purified H44 monoclonal antibody (mAb) was coupled to a 5-mL HiTrap NHS-activated HP column (GE Healthcare). Pooled human blood serum (120 mL) was applied to this affinity column. The IL-18R alpha complex was eluted with elution buffer at a flow rate of 2 mL/minute. Every 1 mL of the elution buffer was collected into a test tube containing 50 mL of neutralization buffer (collected fractions were denoted in order as fractions 1, 2, 3 and so on). A 10-μL aliquot of every fraction (fractions 1 to 8) was treated with the same volume of sample buffer containing 4% SDS (Tris-Glycine SDS Sample Buffer (2×); Invitrogen). Electrophoresis was carried out in the presence of 0.1% SDS, and the gel was stained with Coomassie Brilliant Blue. (B) Western blots of the serum IL-18R alpha complex using an antihuman IL-18R alpha mAb are shown. Western blot analysis was performed using antihuman IL-18R alpha mAb 70625 (R&D Systems, Inc.). Lane 1: Western blot showing 1 μg of rhIL-18R alpha /Fc chimera protein (R&D Systems, Inc.). Lane 2: Western blot showing 5 μg of isolated serum IL-18R alpha complex. (C) Western blot showing serum IL-18R alpha complex using an antihuman IL-18 mAb. Western blot analysis was performed using antihuman IL-18 mAb clone 8 with 5 μg of isolated serum IL-18R alpha complex. (D) Western blot showing serum IL-18R alpha complex using an antihuman IL-18R beta mAb. Western blot analysis was performed using antihuman IL-18R beta mAb 132016 (R&D Systems, Inc.) with 5 μg of isolated serum IL-18R alpha complex. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/21435242), licensed under a CC-BY license. Not internally tested by R&D Systems.