Human IL‑18/IL‑1F4 Propeptide Antibody

Name: Human IL‑18/IL‑1F4 Propeptide Antibody
Brand: R&D Systems
SKU: AF646
Price: 489 USD
Availability: InStock

R&D Systems | Catalog # AF646

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Citations (3)

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Knockout Validated, Western Blot, Simple Western

Cited:

Immunohistochemistry-Paraffin, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all IL-18/IL-1F4 Primary Antibodies »

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 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 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.

Detection of Rhesus Macaque IL-18/IL-1F4 by Immunohistochemistry

CD8+ T- and CD20+ B-lymphocytes are not localized within inflammatory foci.Immunohistochemistry using anti-CD20 identified a few B-lymphocytes in the inflammatory foci (arrows) (A). In contrast, no CD8+ T-lymphocytes are localized using anti-CD8 antibody (B) (400X). Immunohistrochemistry using anti-pro-IL-18 (C) or anti-CXCL9 (D) reveals the expression of these molecules on mononuclear cells within inflammatory foci (200X). Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0014429), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Rhesus Macaque IL-18/IL-1F4 by Immunohistochemistry

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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

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

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.

Calculators

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.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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.