Human IL6 kit
No-wash kit to quantify released Human IL6
This HTRF kit is designed for the quantification of human IL1 beta release in cell supernatant.
IL1 beta, also known as leukocytic pyrogen or mononuclear cell factor, is a pro-inflammatory cytokine of the IL1 family. IL1 beta is considered to be a major endogenous pyrogen and induces prostaglandin synthesis, T cell activation, B cell activation, and subsequent antibody production. It is also a known promoter of T cell differentiation into Th17.
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 IL1 beta analysis.
|Sample size||16 µL|
|Final assay volume||20 µL|
|Kit components||Lyophilized standard, frozen detection antibodies, buffers &protocol.|
|LOD &LOQ (in Diluent)||5 pg/mL &39.4 pg/mL|
|Range||39.4 6,500 pg/mL|
|Time to result||Overnight at RT|
|Calibration||NIBSC (86/680) value (IU/mL) = 0,1 x HTRF hIL1ß value (pg/mL)|
|Sample||Mean [IL1ß] (pg/mL)||CV|
Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.
|Sample||[IL1ß] (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||[IL1β] expected (pg/mL)||[IL1β] detected (pg/mL)||Recovery|
The excellent % of recovery obtained from these experiments show the good linearity of the assay.
|[IL1β] added (pg/mL)||[IL1β] expected (pg/mL)||[IL1β] 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 observed validates the sample matrix used for this assay.
THP1 cells plated at 50 and 100 kcells/well were stimulated for 18 h with PMA and LPS respectively used at 50 ng/mL and 1 µg/mL. 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed with the Human IL1ß Assay.
PBMC plated at 50, 100, 200 and 400 kcells/well were stimulated for 18 h with increasing concentrations of LPS ranging from 0.02 to 2 µg/mL. 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed by the Human IL1ß Assay Kit.
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