Human/Rhesus Macaque IL-18/IL-1F4 Antibody

Name: Human/Rhesus Macaque IL-18/IL-1F4 Antibody
Brand: R&D Systems
SKU: AF2548

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AF2548

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Species Reactivity:	Human, Rhesus Macaque
Immunogen:	E. coli-derived recombinant rhesus macaque IL‑18/IL-1F4 Tyr37-Asp193 Accession # AAK13416
Applications:	Block/Neutralize, ELISA, Immunocytochemistry/Immunofluorescence, Knockout Validated, Simple Western, Western Blot
Label:	Unconjugated
Source:	Polyclonal Goat IgG
Specificity:	Detects rhesus macaque IL‑18/IL-1F4 in direct ELISAs and Western blots. In Western blots, approximately 10% cross-reactivity with recombinant mouse IL-18, recombinant rat IL-18, and recombinant porcine IL-18 is observed.

View Full Technical Details for Human/Rhesus Macaque IL-18/IL-1F4 Antibody

Applications for Human/Rhesus Macaque IL-18/IL-1F4 Antibody

	Recommended Concentration	Sample
Western Blot	0.5 µg/mL	See below
Simple Western	50 µg/mL	See below
ELISA	This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human IL‑18/IL‑1F4 Monoclonal Antibody (Catalog # MAB91242). This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human Total IL-18 DuoSet ELISA Kit (Catalog # ) for convenient development of a sandwich ELISA or the Human Total IL-18/IL-1F4 Quantikine ELISA Kit (Catalog # ) for a complete optimized ELISA.
Immunocytochemistry	5-15 µg/mL	See below

Item	Value
Neutralization	Measured by its ability to neutralize IL‑18/IL‑1F4-induced IFN‑ gamma secretion in the KG‑1 human acute myelogenous leukemia cell line. Novick, D. et al. (1999) Immunity 10(1):127. The Neutralization Dose (ND₅₀) is typically 0.4‑1.2 µg/mL in the presence of 10 ng/mL Recombinant Rhesus Macaque IL‑18/IL‑1F4 and 20 ng/mL Recombinant Human TNF‑ alpha.
Knockout Validated	IL‑18/IL‑1F4 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in IL‑18/IL‑1F4 knockout HeLa cell line.

Please Note: Optimal dilutions should be determined by each laboratory for each application.

Scientific Data Examples for Human/Rhesus Macaque IL-18/IL-1F4 Antibody

	Western Blot Detection of Human IL‑18/IL‑1F4 by Western Blot. Western blot shows lysates of PC-3 human prostate cancer cell line, MCF 10A human breast epithelial cell line, A431 human epithelial carcinoma cell line, and HEK293T human embryonic kidney cell line (negative control cell line). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Rhesus Macaque IL-18/IL-1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for IL-18/IL-1F4 at approximately 22 kDa (as indicated). GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
	Immunocytochemistry IL‑18/IL‑1F4 in Human PBMCs. IL-18/IL-1F4 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human/Rhesus Macaque IL-18/IL-1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (green). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
	Simple Western Detection of Human IL‑18/IL‑1F4 by Simple Western^TM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma parental cell line and IL-18/IL-1F4 knockout HeLa cell line, loaded at 0.2 mg/mL. A specific band was detected for IL-18/IL-1F4 at approximately 29 kDa (as indicated) in the HeLa parental cell line using 50 µg/mL of Goat Anti-Human/Rhesus Macaque IL-18/IL-1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 2-40. kDa separation system.
	Neutralization IFN‑ gamma Secretion Induced by IL‑18/IL‑1F4 and Neutralization by Primate IL‑18/IL‑1F4 Antibody. In the presence of Recombinant Human TNF-alpha (20 ng/mL, Catalog # 210-TA), Recombinant Rhesus Macaque IL-18/IL-1F4 (Catalog # 2548-RM) stimulates 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 (Catalog # DIF50C). Under these conditions, IFN-gamma secretion elicited by Recombinant Rhesus Macaque IL-18/IL-1F4 (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human/Rhesus Macaque IL-18/IL-1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548). The ND₅₀ is typically 0.4-1.2 µg/mL.
	Knockout Validated Western Blot Shows Human IL‑18/IL‑1F4 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and IL-18/IL-1F4 knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Rhesus Macaque IL-18/IL-1F4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2548) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for IL-18/IL-1F4 at approximately 25 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Endotoxin level	<0.10 EU per 1 μg of the antibody by the LAL method.
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
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	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.

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Background: IL-18/IL-1F4

Interleukin-18 (IL-18), also known as IL-1F4 and IFN-gamma inducing factor (IGIF), is a member of the IL-1 family of cytokines and is a key molecule in the innate immune response (1). Rhesus IL-18 is synthesized as a 24 kDa proprotein that contains a 36 amino acid (aa) propeptide and a 157 aa mature region (2). Under inflammatory conditions, the propeptide is cleaved by Caspase-1 in the cytoplasm to liberate the mature nonglycosylated 18 kDa monomeric IL-18 (3, 4). Mature rhesus IL-18 shares 96% aa sequence identity with human IL-18 and 60-76% with mouse, rat, canine, feline, and porcine IL-18. IL-18 is secreted by a variety of cell types including macrophages, dendritic cells, and epithelial cells (1, 5). Circulating mature IL-18 is sequestered by soluble IL-18 binding proteins (IL-18 BP) that inhibit IL-18 bioactivity (6). IL-18 interacts with the widely expressed IL-18 R alpha which then recruits the signaling subunit IL-18 R beta (7, 8). The IL-1 family member IL-1F7 also binds to IL-18 R alpha but does not recruit IL-18 R beta or induce signaling (9). IL-1F7 binds IL-18 BP and enhances its neutralizing effect on IL-18 activity (9). IL-18 synergizes with other cytokines to activate NK, Th1, and Th17 cells and to increase the production of IFN-gamma (1, 5, 10-12). IL-18 can also promote Th2 cytokine release which reduces the effectiveness of antiviral responses (13, 14). Increased levels of active IL-18 contribute to the severity of autoimmunity and hypertension, while deficiency of IL-18 results in symptoms of metabolic syndrome (1, 5, 15, 16). In cancer, IL-18 stimulates Th1 and NK cells to target tumor cells, but it can also promote angiogenesis, metastasis, and tumor cell immune evasion (11).

Item	Value
References	Arend, W.P. et al. (2008) Immunol. Rev. 223:20. Giavedoni, L.D. et al. (2001) J. Interferon Cytokine Res. 21:173. Ghayur, T. et al. (1997) Nature 386:619. Gu, Y. et al. (1997) Science 275:206. Boraschi, D. and C.A. Dinarello (2006) Eur. Cytokine Netw. 17:224. Novick, D. et al. (1999) Immunity 10:127. Torigoe, K. et al. (1997) J. Biol. Chem. 272:25737. Born, T.L. et al. (1998) J. Biol. Chem. 273:29445. Bufler, P. et al. (2002) Proc. Natl. Acad. Sci. USA 99:13723. Takeda, K. et al. (1998) Immunity 8:383. Park, S. et al. (2007) Cell. Mol. Immunol. 4:329. Yoshimoto, T. et al. (1998) J. Immunol. 161:3400. Hoshino, T. et al. (2001) J. Immunol. 166:7014. Iannello, A. et al. (2009) AIDS Rev. 11:115. Rabkin, S.W. (2009) Nat. Clin. Pract. Cardiovasc. Med. 6:192. Netea, M.G. et al. (2006) Nat. Med. 12:650.
Long Name	Interleukin 18
Entrez Gene IDs	3606 (Human); 16173 (Mouse); 29197 (Rat); 397057 (Porcine); 574151 (Primate)
Alternate Names	IFN-gamma-inducing factor, IGIF, IGIFIL-1 gamma, IL-18, IL-18MGC12320, IL-1F4, IL-1g, IL18, IL1F4iboctadekin, Iboctadekin, Interferon gamma-inducing factor, Interleukin-1 gamma, interleukin 18 (interferon-gamma-inducing factor), interleukin-18

Citations for Human/Rhesus Macaque IL-18/IL-1F4 Antibody (1)

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Citations are publications that use Bio-Techne products. Selected citations for Human/Rhesus Macaque IL-18/IL-1F4 Antibody include:

Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

SD Rao et al. (2022), M24B aminopeptidase inhibitors selectively activate the CARD8 inflammasome Nature Chemical Biology, 0(0).
PMID: 35165443

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Product Documents for Human/Rhesus Macaque IL-18/IL-1F4 Antibody

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Human/Rhesus Macaque IL-18/IL-1F4 Antibody

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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Applications for Human/Rhesus Macaque IL-18/IL-1F4 Antibody

Scientific Data Examples for Human/Rhesus Macaque IL-18/IL-1F4 Antibody

Western Blot

Immunocytochemistry

Simple Western

Neutralization

Knockout Validated

Formulation, Preparation and Storage

Reconstitution Calculator

Background: IL-18/IL-1F4

Citations for Human/Rhesus Macaque IL-18/IL-1F4 Antibody (1)

Product Documents for Human/Rhesus Macaque IL-18/IL-1F4 Antibody

Product Datasheets

COA

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