The Human IL18 kit is designed for the quantification of human IL18 release in cell supernatant.
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  • No-wash
  • Low sample consumption
  • Under 2h bench time
The Human IL18 kit is designed for the quantification of human IL18 release in cell supernatant.
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In stock

Overview

Also called IGIF (interferon gamma inducing factor), IL18 is a pro-inflammatory cytokine secreted mainly by macrophages. Its biological activity is close to that of IL12, and it can enhance NK cell activity as well as induce IFNg secretion by PBMC. An important role of IL18 is eliminating VEGF from the eye, thus protecting the retina. It is used together with anti-VEGF antibodies in the treatment of AMD (age-related macular degeneration). In addition to this physiological role, IL18 dysregulation leads to severe inflammatory reactions.

Assay principle

Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection by HTRF® reagents (384-well low-volume white plate or Cisbio low-volume 96-well plate in 20 µl). The antibodies labeled with the HTRF donor and acceptor are pre-mixed and added in a single dispensing step, to further streamline the assay procedure. The assay can be run up to a 1536-well format by simply resizing each addition volume proportionally.

Assay data analysis

The 4 Parameter Logistic (4PL) curve is commonly recommended for fitting an ELISA standard curve. This regression enables the accurate measurement of an unknown sample across a wider range of concentrations than linear analysis, making it ideally suited to the analysis of biological systems like cytokine releases.

To fully understand how to deal with HTRF data processing and also 4PL 1/y² fitting, please visit this page.

Cisbio also worked with Myassays.com to help you in your data analysis. By clicking on this link, you’ll be able to access a free online software to run your IL18 analysis.

Technical specifications of human IL18 kit

Sample size 16 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, buffers &protocol.
LOD &LOQ (in Diluent) 18 pg/mL &43 pg/mL
Range 43 – 8,000 pg/mL
Time to result 1h at RT
Calibration NIBSC (03/200) value (IU/mL) = 5 x HTRF hIL18 value (pg/mL)
Species Human only

Intra and inter assay

Intra-assay (n=24)

Sample Mean [IL18] (pg/mL) CV
1 100 10%
2 4500 3%
3 6800 4%
Mean CV 6%

Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.

Inter-assay (n=4)

Sample [IL18] (pg/mL) Mean (delta R) CV
1 155 410 3%
2 751 1855 5%
3 3636 6570 5%
Mean CV 4%

Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.

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