The HTRF human IL18 kit is designed for the quantification of human IL18 release in cell supernatant.
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).
Feature | Specification |
---|---|
Application | Protein Quantification |
Sample Volume | 16 µL |
The HTRF human IL18 kit is designed for the quantification of human IL18 release in cell supernatant.
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).
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 eye, thus protecting retina. It is used toger with anti-VEGF antibodies in treatment of AMD (age-related macular degeneration). In addition to this physiological role, IL18 dysregulation leads to severe inflammatory reactions.
Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable detection of low levels of analytes in serum or plasma.
Application |
Protein Quantification
|
---|---|
Brand |
HTRF
|
Detection Modality |
HTRF
|
Product Group |
Kit
|
Sample Volume |
16 µL
|
Shipping Conditions |
Shipped in Dry Ice
|
Target Class |
Cytokines
|
Target Species |
Human
|
Technology |
TR-FRET
|
Unit Size |
500 Assay Points
|
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 Revvity 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.
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. Revvity also worked with Myassays.com to help you in your data analysis. you’ll be able to access a free online software to run your IL18 analysis.
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-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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