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Human IL11 kit HTRF®

The HTRF human IL11 kit is designed for the quantification of human IL11 release into cell supernatant.

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The HTRF human IL11 kit is designed for the quantification of human IL11 release into cell supernatant.

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Overview

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.

Benefits

  • SCALABLE FROM 96 TO 1536 WELLS
  • VALIDATED ON MDA-MB 231 CELLS
  • CORRELATED TO ELISA

Assay Principle

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.

HTRF human IL11 assay principle

Assay data analysis

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.

Cisbio has 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 IL11 analysis.

Technical specifications of human IL11 kit

Sample size16 µL
Final assay volume20 µL
Kit componentsLyophilized standard, frozen detection antibodies, buffers, &protocol.
LOD & LOQ (in Diluent) 21 pg/mL &  31 pg/mL
Range31 – 5,000 pg/mL
Time to resultOvernight at RT 
CalibrationNIBSC (92/788) value (UI/ml) = 0.0093 x HTRF hIL11 value (pg/ml)
SpeciesHuman only

Intra and Inter assay



Intra-assay (n=24)

SampleMean [IL11] (pg/mL)CV
178

11

225003.1
350003

Mean CV5.7

Each of the 3 samples was measured 24 times, and the %CV for the concentration was calculated for each sample.

 Inter-assay (n=4)

Sample[IL11] (pg/mL)Mean (delta R)CV
13291311

9

21361440314
35516551614


Mean CV12

Each of the samples was measured in 4 different experiments, and the %CV for the concentration was calculated for each sample.

Spike and recovery

Sample[IL11] recomb prot (pg/mL)[IL11] native prot (pg/mL)% recomb prot% native prot[Il11]expected (pg/mL)[IL11]detected (pg/mL)Recovery
1

2500

67534661295.51467113 %
2250092150501710.5157092%
32500181966342268.5207391%

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.

IL11 secretion of MDA-MB 231 stimulated with TGFb

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.

Validation of HTRF human IL11 kit on MDA-MB 231 cell line

HTRF Product Catalog 2020 July update

All your HTRF assays in one document! - Catalog

Cytokine assays: a guide to success with HTRF

Setting up a successful cytokine assay - Guides

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

An innate and adaptive immunity recap

Insight into the diversity of cells & signaling pathways - Guides

How to measure multiple cytokines from 1 sample using HTRF assays

3' video to set up your cytokine panel - Videos

HTRF makes cytokine detection easy

A fun video introducing you to cytokines assays with HTRF - Videos

How to measure multiple cytokines from 1 sample

What to expect at the bench - Videos

Portfolio of HTRF cytokine assays

This leaflet presents the analytical performances of each assay - Brochures

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Product Insert IL11 (h) Kit / 62HIL11PEG-62HIL11PEH

62HIL11PEG-62HIL11PEH - Product Insert

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

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