Human Total IL-18 Quantikine QuicKit ELISA

Catalog # Availability Size / Price Qty
QK318
Control Products Available
Human IL-18/IL-1F4 ELISA Serum/Plasma Standard Curve
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Human Total IL-18 Quantikine QuicKit ELISA Summary

Assay Length
80 minutes
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL)
Sensitivity
4.57 pg/mL
Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human IL-18.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine® QuicKit™ Human Total IL-18/IL-1F4 Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human IL-18 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-18 and has been shown to accurately quantitate the recombinant factor free and in complex with IL-18 BPa. Results obtained using natural human IL-18 showed linear curves that were parallel to the standard curves obtained using the QuicKit™ standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-18.

Precision

Intra-Assay Precision (Precision within an assay) Two samples of known concentration were tested twenty times on one plate to assess&nbsp;intra-assay precision
Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision. Assays were performed by at least three technicians

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 1 2
n 20 20 10 10
Mean 105 507 114 552
Standard Deviation 4.15 12.8 6.12 43.8
CV% 4 2.5 5.4 7.9

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 1 2
n 20 20 10 10
Mean 120 591 131 638
Standard Deviation 4.23 22 6.9 38.1
CV% 3.5 3.7 5.3 6

Recovery

The recovery of human IL-18 spiked to three different level throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 117 109-128
EDTA Plasma (n=2) 116 108-123
Heparin Plasma (n=2) 101 93-112
Serum (n=2) 101 85-117

Linearity

To assess the linearity of the assay, samples spiked with high concentrations of human IL-18 were serially diluted with the appropriate calibrator diluent to produce samples with values within the dynamic range of the assay.
Human IL-18/IL-1F4 ELISA Linearity

Data Examples

Human IL-18/IL-1F4 ELISA Serum/Plasma Standard Curve

Human IL-18 /IL-1F4 ELISA Cell Culture Supernate Standard Curve

Human Total IL-18 QuicKit Spiked Recovery Competitor Comparison Total IL-18 is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Serum recovery is 77% compared to 61% for the top competitor. EDTA plasma recovery is 89% compared to 60% for the top competitor. Heparin plasma recovery is 88% compared to 65% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.

Human Total IL-18 QuicKit Spiked Linearity Competitor Comparison Total IL-18 is spiked at high concentration in various matrices and diluted with appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. The linearity is between 93%-104% compared to 58%-198% for the top competitor.

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-18/IL-1F4

Interleukin-18 (IL-18) is a proinflammatory cytokine in the IL-1 family that exerts distinct immune effects depending on the local cytokine environment. It is expressed as a 24 kDa precursor by endothelial and epithelial cells, keratinocytes, gamma δT cells, and phagocytes. The precursor is activated intracellularly by Caspase-1 mediated proteolysis to release the 17 kDa mature cytokine. The precursor can also be released by necrotic cells for extracellular cleavage by multiple proteases. IL-18 activation is induced by infection or tissue damage and contributes to disease pathology in chronic inflammation (1-3). IL-18 binds to the widely expressed IL-18 R alpha which recruits IL-18 R beta to form the signaling receptor complex (4, 5). Its bioactivity is negatively regulated by interactions with IL-18 binding proteins and virally encoded IL-18BP homologs (6). In the presence of IL-12 or IL-15, IL-18 enhances anti-viral Th1 immune responses by inducing IFN-gamma production and the cytolytic activity of CD8+ T cells and NK cells (7, 8). In the absence of IL-12 or IL-15, however, IL-18 promotes production of the Th2 cytokines IL-4 and IL-13 by CD4+ T cells and basophils (9, 10). In the presence of IL-1 beta or IL-23, IL-18 induces the antigenindependent production of IL-17 by gamma δ T cells and CD4+ T cells (11). IL-18 also promotes myeloid dendritic cell maturation and triggers neutrophil respiratory burst (12, 13). In cancer, IL-18 exhibits diverse activities including enhancing anti-tumor immunity, inhibiting or promoting angiogenesis, and promoting tumor cell metastasis (14). Mature human IL-18 shares approximately 63% amino acid sequence identity with mouse and rat IL-18 (15). Alternative splicing in human ovarian cancer generates an isoform that is resistant to Caspase-1 activation (16). A cell surface form can be expressed on M-CSF induced macrophages and released in response to bacterial endotoxin (17).

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

Assay Procedure

These assays utilize an anti-tag coated microplate. Standards, samples, and controls are added to plate, followed by subsequent addition of an antibody cocktail. Following a 1 hour incubation, plates are washed and substrate is added and incubated for 20 minutes. Stop solution is then added and plates are read using a standard microplate reader.

QuicKit ELISA assay procedure

FAQs

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