No-wash kit to quantify released Human CCL2 (MCP1)
Human IL6 kit
This HTRF kit is designed for the quantification of IL6 release in cell supernatant.
- Validated on PBMC
- Under 2h bench time
IL6 is a pro-inflammatory cytokine involved in acute-phase reaction, inflammation, and cancer progression. Secreted by T cells, macrophages, and fibroblasts, it induces B and T cell proliferation. It is being studied in a wide variety of research areas including diabetes, Alzheimer's disease, depression, and several cancers. Along with TGF beta, IL6 is the main promoter of T cell differentiation into TH17, a new component of immuno-oncology.
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 IL6 analysis.
|Sample size||16 µL|
|Final assay volume||20 µL|
|Kit components||Lyophilized standard, frozen detection antibodies, buffers &protocol.|
|LOD &LOQ (in Diluent)||8 pg/mL &34 pg/mL|
|Range||34 7,500 pg/mL|
|Time to result||2h at RT|
|Calibration||NIBSC (89/548) value (IU/mL) = 0,1 x HTRF hIL6 value (pg/mL)|
|Sample||Mean [IL6] (pg/mL)||CV|
Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.
|Sample||[IL6] (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||[IL6] expected (pg/mL)||[IL6] detected (pg/mL)||Recovery|
The excellent % of recovery obtained from these experiments show the good linearity of the assay.
|[IL6] added (pg/mL)||[IL6] expected (pg/mL)||[IL6] detected (pg/mL)||Recovery|
Recombinant cytokine was added to a serum sample, and the response obtained from a standard curve was compared to the calculated expected values. The 100% of recovery observed validates the sample matrix used for this assay.
THP1 cells plated at 100 kcells/well were incubated for 18 h with increasing concentrations of JTE 607 in presence of 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 IL6 Assay.
PBMC were plated at 50, 100 and 400 kcells/well and stimulated for 18 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 IL6 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 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
cGAS-STING signaling pathway from A to Z - Brochures
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