Human IL-18 BPa Antibody
R&D Systems, part of Bio-Techne | Catalog # AF119
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Cited:
Human, Mouse
Applications
Validated:
ELISA, Neutralization, Western Blot
Cited:
ELISA Development, Neutralization, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant human IL-18 BPa
Thr31-Gly194
Accession # O95998
Thr31-Gly194
Accession # O95998
Specificity
Detects human IL‑18 BPa in direct ELISAs and Western blots.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Human IL-18 BPa Antibody
Detection of Human IL‑18 BPa by Western Blot.
Western blot shows conditioned media from HUVEC human umbilical vein endothelial cells untreated (-) or treated (+) with 50 ng/mL Recombinant Human IFN-gamma (Catalog # 285-IF) and 40 ng/mL Recombinant Human TNF-alpha (210-TA) for 48 hours. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human IL-18 BPa Antigen Affinity-purified Polyclonal Antibody (Catalog # AF119) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for IL-18 BPa at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
IL‑18 BPa Inhibition of IL‑18/ IL‑1F4-induced IFN‑ gamma Secretion and Neutralization by Human IL‑18 BPa Antibody.
Recombinant Human IL-18 BPa Fc Chimera (119-BP) inhibits Recombinant Human IL-18/IL-1F4 induced 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 (DIF50C). Inhibition of Recombinant Human IL-18/IL-1F4 (40 ng/mL) activity elicited by Recombinant Human IL-18 BPa Fc Chimera (0.1 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL-18 BPa Antigen Affinity-purified Polyclonal Antibody (Catalog # AF119). The ND50 is typically 1-5 µg/mL in the presence of Recombinant Human TNF-alpha (20 ng/mL).
Human IL‑18/IL-18 BPa Complex ELISA Standard Curve.
Recombinant Human IL-18/IL-18 BPa complex protein was serially diluted 2-fold and captured by Rabbit Anti-Human IL-18/IL-1F4 Monoclonal Antibody (Catalog # MAB91244) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human IL-18 BPa Antigen Affinity-purified Polyclonal Antibody (Catalog # AF119) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).Applications for Human IL-18 BPa Antibody
Application
Recommended Usage
ELISA
This antibody functions as an ELISA detection antibody when paired with Rabbit Anti-Human IL‑18/IL‑1F4 Monoclonal Antibody (Catalog # MAB91244).
This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human IL-18/IL-18 BPa Complex DuoSet ELISA Kit (Catalog # DY8936-05) for convenient development of a sandwich ELISA or the Human IL-18 BPa Quantikine ELISA Kit (Catalog # DBP180) for a complete optimized ELISA.
Western Blot
0.5 µg/mL
Sample: HUVEC human umbilical vein endothelial cells treated with Recombinant Human IFN‑ gamma (Catalog # 285-IF) and Recombinant Human TNF‑ alpha (Catalog # 210-TA)
Sample: HUVEC human umbilical vein endothelial cells treated with Recombinant Human IFN‑ gamma (Catalog # 285-IF) and Recombinant Human TNF‑ alpha (Catalog # 210-TA)
Neutralization
Measured by its ability to neutralize IL‑18 BPa inhibition of IL‑18/IL‑1F4-induced IFN‑ gamma secretion in the KG‑1 human acute myelogenous leukemia cell line. The Neutralization Dose (ND50) is typically 1-5 µg/mL in the presence of 0.1 µg/mL Recombinant Human IL‑18 BPa Fc Chimera, 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.
Reviewed Applications
Read 2 reviews rated 4.5 using AF119 in the following applications:
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 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. *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 BPa
Interleukin 18 binding protein (IL-18 BP) is a secreted glycoprotein, which functions as an IL-18 antagonist by binding to IL-18 and blocking its biological activity. IL-18 BP bears no amino acid sequence homology to the membrane-associated IL-18 and IL-1 receptor proteins. The gene for human IL-18 BP has been localized to chromosome 11q13. It encodes for at least four isoforms by alternative splicing. The IL-18 BP isoforms a and c each contain one immunoglobulin (Ig)-like C2-type domain while isoforms b and d lack a complete Ig domain. The complete Ig domain has been shown to be essential to the binding and neutralizing properties of the binding proteins. Two isoforms of mouse IL18 BP (c and d) containing the complete Ig domain have also been isolated and shown to neutralize IL-18 bioactivity. Human and mouse IL-18 BPs share approximately 61% amino acid sequence identity. Several poxviruses also encode proteins with sequence similarity to the human and mouse IL-18 BP. Viral IL-18 BPs have been shown to bind and inhibit IL-18 responses and may be involved in modulating host immune responses. The expression of IL-18 BP is markedly up-regulated by IFN-gamma, suggesting that IL-18 activity is modulated by a negative feedback mechanism mediated by IL-18 BP.
References
- Mühl, H. et al. (2000) Biochem. Biophys. Res. Commun. 267:960.
- Kim, S-H. et al. (2000) Proc. Nat. Acad. Sci. USA 97:1190.
- Calderara, S. et al. (2001) Virology 279:22.
Long Name
Interleukin 18 Binding Protein Isoform a
Alternate Names
IL18 BPa, IL18BP
Gene Symbol
IL18BP
UniProt
Additional IL-18 BPa Products
Product Documents for Human IL-18 BPa Antibody
Product Specific Notices for Human IL-18 BPa Antibody
For research use only
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