No-wash kit to quantify released Human CCL2 (MCP1)
Human IL8 kit
This HTRF kit is designed for the quantification of human IL8 release in cell supernatant.
- Validated on PBMC
- Under 2h bench time
Also called CXCL8, IL8 is a chemokine mainly produced by macrophages, T cells, and neutrophils. IL8 acts as a chemoattractant for neutrophils, and promotes infiltration and activation at inflammation sites. IL8 is involved in several cancer types due to its ability to promote angiogenesis and cell proliferation, as well as to inhibit apoptosis.
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 IL8 analysis.
|Sample size||16 µL|
|Final assay volume||20 µL|
|Kit components||Lyophilized standard, frozen detection antibodies, buffers &protocol.|
|LOD &LOQ (in Diluent)||6 pg/mL &32 pg/mL|
|Range||32 4,000 pg/mL|
|Time to result||1h at RT|
|Calibration||NIBSC (89/520) value (IU/mL) = 0,01 x HTRF hIL8 value (pg/mL)|
|Sample||Mean [IL8] (pg/mL)||CV|
Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.
|Sample||[IL8] (pg/mL)||Mean (delta R)||CV|
Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.
|Dilution factor||[IL8] expected (pg/mL)||[IL8] detected (pg/mL)||Recovery|
The excellent % of recovery obtained from these experiments show the good linearity of the assay.
|[IL8] added (pg/mL)||[IL8] expected (pg/mL)||[IL8] detected (pg/mL)||Recovery|
The same amount of recombinant cytokine was added to 2 different serum samples, and the set of responses obtained from a standard curve was compared to the calculated expected values. The ~ 100% of recovery found validates the sample matrix used for this assay.
THP1 cells plated at 100 kcells/well were incubated with increasing concentrations of JTE 607 for 18h, then stimulated for 3 h with 2 µg/mL LPS. 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed by the Human IL8 Assay.
PBMC plated at 50, 100, 200, and 400 kcells/well were stimulated for 3 h with increasing concentrations of LPS (0, 0.02, 0.2, 2 µg/mL). 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed by the Human IL8 Assay.
Can HTRF technology answer to different scientific questions? An academic with industrial standards point of view
In collaboration with CNRS - Scientific Presentations
In collaboration with GSK - Scientific Presentations
In collaboration with Genomics institute of the Novartis Research Foundation - Scientific Presentations
In collaboration with Molecular Devices - Application Notes
In collaboration with Blood assay solution - Application Notes
Perform and optimize cytokine assays on PBMC - Technical Notes
A fun video introducing you to cytokines assays with HTRF - Videos
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