Human IL-18 R beta/IL-1 R7 Antibody (132016)

IFN-gamma secretion Induced by IL-18/IL1F4 and Neutralization by Human IL-18 R beta /IL-1 R7 Antibody.

In the presence of Recombinant Human TNF-alpha (20 ng/mL, Catalog # 210-TA), Recombinant Human IL-18/ IL-1F4 stimulates IFN-gamma secretion in the KG-1 human acute myelogenous leukemia cell line in a dose-dependent manner (left-hand graph), 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 (right-hand graph) by increasing concentrations of Mouse Anti-Human IL-18 R beta /IL-1 R7 Monoclonal Antibody (Catalog # MAB1181). The ND₅₀ is typically 0.3-1.0 µg/mL.

Detection of Human IL-18 R beta/IL-1 R7/ACPL 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.