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Human Total IL-18/IL-1F4 Quantikine ELISA Kit

R&D Systems | Catalog # DL180

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Key Product Details

Assay Length

4.0 hours

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)

Sensitivity

5.15 pg/mL

Assay Range

15.6-1000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
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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)

Conjugate

HRP

Species

Human

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 TypeMean (pg/mL)Range (pg/mL)Standard Devitation (pg/mL)
Serum (n=30)18384.8-35869.7
EDTA plasma (n=30)18381.5-34468.1
Heparin plasma18287.5-33365.8

Sample TypeMean of Detectable (pg/mL)% DetectableRange (pg/mL)
Saliva (n=10)57.170ND-69.8
Urine (n=10)44.030ND-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)
Unstimulated19.6
Stimulated562





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.

Human Total IL-18 ELISA Linearity

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 Cell Culture Supernate/Urine Standard Curve

Human Total IL-18 ELISA Serum/Plasma/Saliva 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 delta T 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).

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

Additional IL-18/IL-1F4 Products

Product Documents for Human Total IL-18/IL-1F4 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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.

Citations for Human Total IL-18/IL-1F4 Quantikine ELISA Kit

Customer Reviews for Human Total IL-18/IL-1F4 Quantikine ELISA Kit (4)

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  • Human Total IL-18/IL-1F4 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 02/25/2023
  • Human Total IL-18/IL-1F4 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 01/27/2021
    Curve developed beautifully, kit is easy to use.
    Human Total IL-18/IL-1F4 Quantikine ELISA Kit DL180
  • Human Total IL-18/IL-1F4 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 01/27/2021
    Picture of the plate before adding the stop solution. Standard curve in duplicate in columns 1 and 2. Assay ran in duplicate. Clear, precise and easy to follow directions.
    Human Total IL-18/IL-1F4 Quantikine ELISA Kit DL180
  • Human Total IL-18/IL-1F4 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 01/25/2020

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FAQs for Human Total IL-18/IL-1F4 Quantikine ELISA Kit

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  • Q: Does the Human Total IL-18/IL-1F4 Quantikine ELISA Kit recognize both free and bound IL-18?

    A: The Human Total IL-18/IL-1F4 Quantikine ELISA Kit detects free and complexed Human IL-18 in validated sample types.

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