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HTRF Human IL-11 Detection Kit, 500 Assay Points

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

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).
Product Variants
Part number: 62HIL11PEG
Unit Size: 500 Assay Points
List price: USD 1,103.00
Your price:
USD 1,103.00
USD 1,103.00 /each
Part number: 62HIL11PEH
Unit Size: 10,000 Assay Points
List price: USD 10,290.00
Your price:
USD 0.00
USD 10,290.00 /each

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.

Specifications

Assay Points
500
Assay Target Type
Kit
Assay Technology
HTRF
Brand
HTRF
Quantity
1
Therapeutic Area
Inflammation
NASH/Fibrosis
Unit Size
500 Assay Points

Video gallery

How it works

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 Revvity 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.

cytokines-how-it-works-assay-principle-human
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.

Revvity has also worked with Myassays.com to help you in your data analysis.

 

Assay details

Technical specifications of human IL11 kit
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)
Species     Human only

 

Analytical performance

Intra and Inter assay

Intra-assay (n=24)

Sample Mean [IL11] (pg/mL) CV
1 78 11
2 2500 3.1
3 5000 3
  Mean CV 5.7

 

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
1 329 1311 9
2 1361 4404 14
3 5516 5516 14
    Mean CV 12

 

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 675 34 66 1295.5 1467 113%
2 2500 921 50 50 1710.5 1570 92%
3 2500 1819 66 34 2268.5 2073 91%

 

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.

 

Assay validation

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.

assay-validation-human

 

Resources

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Guide
HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.

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SDS, COAs, Manuals and more

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