Human IL11 kit HTRF®

Name: Human IL11 kit
Brand: Cisbio
SKU: 44644
Availability: InStock

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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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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.

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 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

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	4403	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.

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

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