Human IL-18/IL-1F4 Propeptide Antibody

Catalog # Availability Size / Price Qty
MAB646
MAB646-SP
Detection of Human IL‑18/IL‑1F4 by Western Blot.
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Product Details
Citations (3)
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Human IL-18/IL-1F4 Propeptide Antibody Summary

Species Reactivity
Human
Specificity
Detects the pro region of human IL-18/IL-1F4 in direct ELISAs and Western blots. Does not cross‑react with recombinant human IL-18, recombinant mouse IL-18 or recombinant rat IL-18.
Source
Monoclonal Mouse IgG1 Clone # 74801
Purification
Protein A or G purified from ascites
Immunogen
E. coli-derived recombinant human IL‑18/IL‑1F4
Accession # Q14116
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Intracellular Staining by Flow Cytometry
0.25 µg/106 cells
See below
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. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot Detection of Human IL-18/IL-1F4 antibody by Western Blot. View Larger

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 2 µg/mL of Mouse Anti-Human IL-18/IL-1F4 Propeptide Monoclonal Antibody (Catalog # MAB646) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for IL-18/IL-1F4 at approximately 22 kDa (as indicated). GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Intracellular Staining by Flow Cytometry Detection of IL-18/IL-1F4 antibody in Human PBMCs antibody by Flow Cytometry. View Larger

Detection of IL‑18/IL‑1F4 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) either untreated (upper panel) or treated with LPS for 24 hours (lower panel) were stained with Mouse Anti-Human IL-18/IL-1F4 Propeptide Monoclonal Antibody (Catalog # MAB646, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin. View our protocol for Staining Intracellular Molecules.

Intracellular Staining by Flow Cytometry Detection of IL-18/IL-1F4 Propeptide antibody in Human THP-1 cells antibody by Flow Cytometry. View Larger

Detection of IL-18/IL-1F4 Propeptide in Human THP-1 cells by Flow Cytometry. THP-1 human acute monocytic leukemia cells were stained with Mouse Anti-Human IL-18/IL-1F4 Propeptide Monoclonal Antibody (Catalog # MAB646, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram) followed by PE-conjugated anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Knockout Validated Western Blot Shows Human IL-18/IL-1F4 Antibody Specificity by Using Knockout Cell Line. View Larger

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 2 µg/mL of Mouse Anti-Human IL-18/IL-1F4 Propeptide Monoclonal Antibody (Catalog # MAB646) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for IL-18/IL-1F4 at approximately 22 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH ) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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.

Background: IL-18/IL-1F4

Pro-IL-18 (pro-Interleukin 18; also pro-IGIF and pro-IL-1 gamma ) is a 24 kDa member of the IL-1 family of molecules. It is widely expressed, being produced by keratinocytes, intestinal epithelium, T cells, macrophages and osteoblasts. Human Pro-IL-18 is 193 amino acids (aa) in length. Although mature IL-18 induces IFN-gamma secretion by NK and T cells, Pro-IL-18 appears to have little intrinsic activity. Generally, active IL-18 is considered to arise from caspase-1 cleavage of Pro-IL-18 between Asp36-Tyr37. This generates an 18 kDa mature C-terminal fragment, and a 4 kDa (predicted) N-terminal prosegment that runs at 6 kDa in SDS-PAGE. Other proteases are known to process Pro-IL-18. Caspase-3 cleavage after Asp68 generates an inactive 14 kDa mature segment, Merpin beta -subunit cleavage after Asn52 generates a marginally active 17 kDa mature segment, while parasite Cys protease cleavage after Val47 generates an inactive 17 kDa mature molecule. One splice variant shows a deletion of aa 27-30. Over aa 2-36, human Pro-IL-18 shares 63% aa identity with mouse Pro-IL-18.

Long Name
Interleukin 18
Entrez Gene IDs
3606 (Human); 16173 (Mouse); 29197 (Rat); 397057 (Porcine); 574151 (Primate)
Alternate Names
Iboctadekin; IFN-gamma-inducing factor; IGIF; IGIFIL-1 gamma; IL18; IL-18; IL-18MGC12320; IL-1F4; IL1F4iboctadekin; IL-1g; Interferon gamma-inducing factor; interleukin 18 (interferon-gamma-inducing factor); Interleukin-1 gamma; interleukin-18

Product Datasheets

Citations for Human IL-18/IL-1F4 Propeptide Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. NLRP3 inflammasome: a novel link between lipoproteins and atherosclerosis
    Arch Med Sci, 2016;12(5):950-958.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  2. IL-18 Is Involved in Eosinophil-Mediated Tumoricidal Activity against a Colon Carcinoma Cell Line by Upregulating LFA-1 and ICAM-1.
    Authors: Gatault S, Delbeke M, Driss V, Sarazin A, Dendooven A, Kahn J, Lefevre G, Capron M
    J Immunol, 2015;195(5):2483-92.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. IL-8 dictates glycosaminoglycan binding and stability of IL-18 in cystic fibrosis.
    Authors: Reeves EP, Williamson M, Byrne B, Bergin DA, Smith SG, Greally P, O'Kennedy R, O'Neill SJ, McElvaney NG
    J. Immunol., 2010;184(3):1642-52.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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