Human IL-18/IL-1F4 Propeptide Antibody
R&D Systems, part of Bio-Techne | Catalog # AF646
Key Product Details
Validated by
Knockout/Knockdown
Species Reactivity
Validated:
Human
Cited:
Human, Primate - Macaca mulatta (Rhesus Macaque)
Applications
Validated:
Knockout Validated, Simple Western, Western Blot
Cited:
Immunocytochemistry, Immunohistochemistry-Paraffin
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
Product Specifications
Immunogen
E. coli-derived recombinant human IL‑18/IL‑1F4
Specificity
Detects the pro region of human IL‑18/IL‑1F4 in direct ELISAs and Western blots. In these formats, approximately 5% cross‑reactivity with mature recombinant human IL-18 is observed and less than 1% cross‑reactivity with mature recombinant mouse IL-18 and recombinant rat IL‑18 is observed.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
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 0.5 µg/mL of Goat Anti-Human IL-18/IL-1F4 Propeptide Antigen Affinity-purified Polyclonal Antibody (Catalog # AF646) 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.
Detection of Human IL‑18/IL‑1F4 by Simple WesternTM.
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 IL-18/IL-1F4 Propeptide Antigen Affinity-purified Polyclonal Antibody (Catalog # AF646) 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 12-230 kDa separation system.
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 0.5 µg/mL of Goat Anti-Human IL-18/IL-1F4 Propeptide Antigen Affinity-purified Polyclonal Antibody (Catalog # AF646) 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) in the parental HeLac ell 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.Applications for Human IL-18/IL-1F4 Propeptide Antibody
Application
Recommended Usage
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.
Simple Western
50 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line
Sample: HeLa human cervical epithelial carcinoma cell line
Western Blot
0.5 µg/mL
Sample: PC‑3 human prostate cancer cell line, MCF 10A human breast epithelial cell line, and A431 human epithelial carcinoma cell line
Sample: PC‑3 human prostate cancer cell line, MCF 10A human breast epithelial cell line, and A431 human epithelial carcinoma cell line
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/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
Gene Symbol
IL18
Additional IL-18/IL-1F4 Products
Product Documents for Human IL-18/IL-1F4 Propeptide Antibody
Product Specific Notices for Human IL-18/IL-1F4 Propeptide Antibody
For research use only
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