Detects Recombinant Human IL-18 R alpha/IL-1 R5 in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant human (rh) IL-1 RI, rhIL-1 RII, rhIL-1 R3, rhIL-1 R4, rhIL-1 R6, rhIL-1 R7, rhIL-1 R8, rhIL-1 R9, rhSIGIRR or recombinant mouse IL-18 R alpha is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for 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.

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Applications for Human IL-18 R alpha/IL-1 R5 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/10⁶ cells
Sample: Human peripheral blood mononuclear cells treated with PHA and Recombinant Human IL-2 (Catalog # 202-IL)

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs)

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human skin

Neutralization

Measured by its ability to neutralize IL‑18/IL‑1F4-induced IFN‑ gamma secretion in KG-1 human myeloid leukemia cells. The Neutralization Dose (ND₅₀) is typically 0.03-0.1 µg/mL in the presence of 40 ng/mL Recombinant Human IL‑18/IL‑1F4 and 20 ng/mL Recombinant Human TNF-alpha.

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: IL-18 R alpha/IL-1 R5

Interleukin 18 (IL-18) is a member of the IL-1 family of cytokines and shares numerous immunoregulatory functions with IL-12. The functional IL-18 receptor complex is composed of two subunits designated IL-18 R alpha (also termed IL-1 R5 and IL-1 Rrp) and IL-18 R beta (also termed IL-1 R7 and AcPL). Both IL-18 R alpha and IL-18 R beta belong to the IL-1 receptor superfamily. Although IL-18 R by itself binds IL-18 with low affinity and IL-18 R beta does not bind IL-18 in vitro, co-expression of IL‑18 R alpha and IL‑18 R beta is required for high affinity binding and IL-18 responsiveness. Human IL-18 R cDNA encodes a 541 amino acid (aa) precursor type I membrane protein with a hydrophobic signal, an extracellular domain comprised of three immunoglobulin-like domains, a transmembrane domain and a cytoplasmic region of approximately 200 aa. Human and mouse IL-18 R share 65% amino acid sequence homology. IL-18 R is widely expressed in numerous tissues including spleen, thymus, leukocyte, liver, lung, heart, small and large intestine, prostate and placenta.

References

Parnet, P. et al. (1996) J. Biol. Chem. 271:3967.
Torigoe, K. et al. (1997) J. Biol. Chem. 272:25737.
Born, T.L. et al. (1998) J. Biol. Chem. 273:29445.

Long Name

Interleukin 18 Receptor alpha

Alternate Names

CD218a, IL-1 R5, IL-1 Rrp, IL-18 R1, IL-18Ra, IL18R alpha, IL18R1

Entrez Gene IDs

8809 (Human); 16182 (Mouse)

Gene Symbol

IL18R1

UniProt

Q13478

Additional IL-18 R alpha/IL-1 R5 Products

Product Documents for Human IL-18 R alpha/IL-1 R5 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human IL-18 R alpha/IL-1 R5 Antibody

For research use only

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Flow Cytometry