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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Scientific Data Images for Human IL-18/IL-1F4 Propeptide Antibody

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.

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.

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.

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Applications for Human IL-18/IL-1F4 Propeptide 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.

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: Human peripheral blood mononuclear cells (PBMCs) treated with LPS or THP‑1 human acute monocytic leukemia cells were fixed with paraformaldehyde and permeabilized with saponin

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.

Western Blot

2 µg/mL
Sample: PC‑3 human prostate cancer cell line, MCF 10A human breast epithelial cell line, and A431 human epithelial carcinoma cell line

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

Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

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. *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/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

Alternate Names

IGIF, IL-1F4, IL-1g, IL18

Entrez Gene IDs

3606 (Human); 16173 (Mouse); 29197 (Rat); 397057 (Porcine); 574151 (Primate)

Gene Symbol

IL18

UniProt

Q14116

Additional IL-18/IL-1F4 Products

Product Documents for Human IL-18/IL-1F4 Propeptide Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human IL-18/IL-1F4 Propeptide Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human IL-18/IL-1F4 Propeptide Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB646

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications