Key Product Details
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (17 µL), EDTA Plasma (17 µL), Heparin Plasma (17 µL), Saliva (25 µL), Urine (50 µL)
Assay Range
15.6-1000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
Product Summary
for Human Total IL-18/IL-1F4 Quantikine ELISA Kit
The Quantikine® Human Total IL-18/IL-1F4 Immunoassay is a 4.0 hour solid-phase ELISA designed to measure human IL-18 in cell culture supernates, serum, plasma, saliva, and urine. 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 Quantikine® kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-18.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Specificity
Natural and recombinant human Total IL-18.
Cross-reactivity
Cross-reactivity observed with 1 or more available related molecules. Cross-species reactivity observed with 1 or more species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma/Saliva/Urine - Samples from apparently healthy volunteers were evaluated for
the presence of human Total IL-18 in this assay. No medical histories were available for the
donors used in this study.
| Sample Type | Mean (pg/mL) | Range (pg/mL) | Standard Devitation (pg/mL) |
| Serum (n=30) | 183 | 84.8-358 | 69.7 |
| EDTA plasma (n=30) | 183 | 81.5-344 | 68.1 |
| Heparin plasma | 182 | 87.5-333 | 65.8 |
| Sample Type | Mean of Detectable (pg/mL) | % Detectable | Range (pg/mL) |
| Saliva (n=10) | 57.1 | 70 | ND-69.8 |
| Urine (n=10) | 44.0 | 30 | ND-74.0 |
ND=Non-detectable
Cell Culture Supernates - THP-1 human acute monocytic leukemia cells were cultured in RPMI
supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 1 μg/mL
LPS for 5 hours followed by 20 μM Nigericin for 1 hour. Aliquots of the cell culture supernates
were removed and assayed for levels of human Total IL-18.
| Condition | (pg/mL) |
| Unstimulated | 19.6 |
| Stimulated | 562 |
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Urine
|
Intra-Assay Precision |
Inter-Assay Precision |
| Sample |
1 |
2 |
3 |
1 |
2 |
3 |
| n |
20 |
20 |
20 |
20 |
20 |
20 |
| Mean (pg/mL) |
175 |
301 |
636 |
197 |
381 |
739 |
| Standard Deviation |
5.49 |
8.06 |
24.6 |
27.4 |
46.7 |
81.1 |
| CV% |
3.1 |
2.7 |
3.9 |
13.9 |
12.3 |
11.0 |
EDTA Plasma, Heparin Plasma, Saliva, Serum
|
Intra-Assay Precision |
Inter-Assay Precision |
| Sample |
1 |
2 |
3 |
1 |
2 |
3 |
| n |
20 |
20 |
20 |
20 |
20 |
20 |
| Mean (pg/mL) |
126 |
251 |
495 |
126 |
248 |
494 |
| Standard Deviation |
3.85 |
6.33 |
14.6 |
10.9 |
19.7 |
41.6 |
| CV% |
3.1 |
2.5 |
2.9 |
8.7 |
7.9 |
8.4 |
Recovery
for Human Total IL-18/IL-1F4 Quantikine ELISA Kit
The recovery of human Total IL-18 spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type |
Average % Recovery |
Range % |
| Cell Culture Media (n=4) |
96 |
90-102 |
| EDTA Plasma (n=4) |
104 |
93-113 |
| Heparin Plasma (n=4) |
102 |
88-110 |
| Saliva (n=4) |
108 |
94-120 |
| Serum (n=4) |
100 |
96-105 |
| Urine (n=4) |
91 |
84-100 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Total IL-18 were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images
for Human Total IL-18/IL-1F4 Quantikine ELISA Kit
Human Total IL-18 ELISA Cell Culture Supernate/Urine Standard Curve
Human Total IL-18 ELISA Serum/Plasma/Saliva Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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 deltaT 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 antigen independent production of IL-17 by gamma delta 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).
Alternate Names
IGIF, IL-1F4, IL-1g, IL18
Additional IL-18/IL-1F4 Products
Product Documents for Human Total IL-18/IL-1F4 Quantikine ELISA Kit
Product Specific Notices
for Human Total IL-18/IL-1F4 Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to
www.P65Warnings.ca.gov.