Human IL11 kit HTRF®
The HTRF human IL11 kit is designed for the quantification of human IL11 release into cell supernatant.
IL11, a member of the IL6 family of cytokines, regulates inflammatory responses and depending on the inflammatory microenvironnement, participates in cardiovascular diseases, cancers, or lung fibrosis.
IL11 is produced by a variety of stromal cells, including fibroblasts, epithelial cells, and osteoblasts, and has a variety of effects on hematopoiesis, on inhibition of adipocytogenesis, osteoclast differentiation, stimulating tissue fibrosis, minimizing tissue injury, and regulating the function of B cells.
After its secretion, IL11 binds to its receptor, which is composed of the IL11Ra chain associated with gp130 co-receptor, and triggers the jak/STAT signaling pathway.
HTRF human IL11 kit is a homogeneous add-and-read assay to measure IL11 secretion in cell supernatants. It offers a fast and efficient alternative to ELISA.
- SCALABLE FROM 96 TO 1536 WELLS
- VALIDATED ON MDA-MB 231 CELLS
- CORRELATED TO ELISA
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.
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 release.
To fully understand how to deal with HTRF data processing, and also 4PL 1/y² fitting, please visit this page.
|Sample size||16 µL|
|Final assay volume||20 µL|
|Kit components||Lyophilized standard, frozen detection antibodies, buffers, &protocol.|
|LOD & LOQ (in Diluent)||21 pg/mL & 31 pg/mL|
|Range||31 – 5,000 pg/mL|
|Time to result||Overnight at RT|
|Calibration||NIBSC (92/788) value (UI/ml) = 0.0093 x HTRF hIL11 value (pg/ml)|
|Sample||Mean [IL11] (pg/mL)||CV|
Each of the 3 samples was measured 24 times, and the %CV for the concentration was calculated for each sample.
|Sample||[IL11] (pg/mL)||Mean (delta R)||CV|
Each of the samples was measured in 4 different experiments, and the %CV for the concentration was calculated for each sample.
|Sample||[IL11] recomb prot (pg/mL)||[IL11] native prot (pg/mL)||% recomb prot||% native prot||[Il11]expected (pg/mL)||[IL11]detected (pg/mL)||Recovery|
Different amounts of recombinant IL11 cytokine were added to L15 Glutamax+ 1% BSA cell supernatant samples containing a known concentration of IL11 . Samples were mixed at a 1:1 volume ratio. The set of responses obtained from a standard curve was compared to the calculated expected values. The recovery value observed validates the sample matrix used for this assay.
Human MDA-MB 231 cells were plated at 400,000 cells/well in a 96-well plate and cultured overnight under 100 µL of complete L15 Glutamax cell culture medium, 37°C, at atmospheric CO2 concentration. The cells were then stimulated with increasing concentrations of TGFb (0,05 to 100 µg/mL) prepared in 200µl of 1% BSA L15 Glutamax medium. After 24h of stimulation, 16 µL of supernatant were transferred into a white detection plate to be analyzed with the Human IL11 assay. The HTRF signal was recorded after an overnight incubation.
Insight into the diversity of cells & signaling pathways - Guides
3' video to set up your cytokine panel - Videos
A fun video introducing you to cytokines assays with HTRF - Videos
This leaflet presents the analytical performances of each assay - Brochures
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