Human IL6 kit HTRF®

The HTRF human IL6 kit is designed for the quantification of IL6 release in cell supernatant.

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  • No-wash No-wash
  • Validated on PBMC Validated on PBMC
  • Under 2h bench time Under 2h bench time

The HTRF human IL6 kit is designed for the quantification of IL6 release in cell supernatant.



IL6 is a pro-inflammatory cytokine involved in acute-phase reaction, inflammation, and cancer progression. Secreted by T cells, macrophages, and fibroblasts, it induces B and T cell proliferation. It is being studied in a wide variety of research areas including diabetes, Alzheimer's disease, depression, and several cancers. Along with TGF beta, IL6 is the main promoter of T cell differentiation into TH17, a new component of immuno-oncology.

Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable the detection of low levels of analytes in serum or plasma. Check out the Human IL-6 AlphaLISA kit and the Human IL-6 AlphaLISA biotin-free kit ’s analytical performances for more information or learn more about AlphaLISA. When assaying, always follow the recommended protocol and avoid highly haemolyzed samples.



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.
IL6 kit 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 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 IL6 analysis.

Technical specifications of human IL6 kit

Sample size 16 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, buffers &protocol.
LOD &LOQ (in Diluent) 8 pg/mL &34 pg/mL
Range 34 – 7,500 pg/mL
Time to result 2h at RT
Calibration NIBSC (89/548) value (IU/mL) = 0,1 x HTRF hIL6 value (pg/mL)
Species Human only

Intra and inter assay

Intra-assay (n=24)

Sample Mean [IL6] (pg/mL) CV
1 384 5%
2 2522 3%
3 8594 3%
Mean CV 4%

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

Inter-assay (n=4)

Sample [IL6] (pg/mL) Mean (delta R) CV
1 116.5 400 13%
2 268 1805 10%
3 3261 7834 2%
Mean CV 8%

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

Dilutional linearity

Dilution factor[IL6] expected (pg/mL)[IL6] detected (pg/mL)Recovery

​The excellent % of recovery obtained from these experiments show the good linearity of the assay.

Spike and recovery

[IL6] added (pg/mL)[IL6] expected (pg/mL)[IL6] detected (pg/mL)Recovery

​Recombinant cytokine was added to a serum sample, and the response 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.

Inhibition of IL6 secretion in THP1 cells with JTE607

THP1 cells plated at 100 kcells/well were incubated for 18 h with increasing concentrations of JTE 607 in presence of 2 µg/mL LPS. 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed by the Human IL6 Assay.

Inhibition of IL6 secretion in THP1 cells with JTE607.

IL6 secretion in PBMCs stimulated with LPS

PBMC were plated at 50, 100 and 400 kcells/well and stimulated for 18 h with increasing concentrations of LPS ( 0, 0.02, 0.2, 2 µg/mL). 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed by the Human IL6 Assay.

IL6 secretion in PBMCs stimulated with LPS.

Identification of synoviocyte activation inhibitors by parallel biomarker profiling.

In collaboration with Genomics institute of the Novartis Research Foundation - Scientific Presentations

HTRF Cytokine cell-based assays

Monitor immune regulation the simple way - Flyers

Biomarker and Cell Signaling Assays for Fibrosis and NASH

HTRF and Alpha solutions for NASH - Flyers

Inflammation cell by cell

HTRF solutions for each cell type - Flyers

HTRF assays for Oncology and Inflammation

Signaling in the Immune System - Brochures

HTRF measurement of cytokine release from fresh blood samples

In collaboration with Blood assay solution - Application Notes

Cytokine secretion profile on PBMCs

One sample to measure them all - Posters

Guidelines from PBMC isolation to cytokine assay optimisation

Perform and optimize cytokine assays on PBMC - Technical Notes

How to freeze PBMC

Guideline for PBMC freezing - Technical Notes

Webinar: NASH - Fibrosis Research and Drug Discovery

Featuring a panel of experts - Videos

How to measure multiple cytokines from 1 sample

What to expect at the bench - Videos

HTRF makes cytokine detection easy

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

STING HTRF offer to bridge innate and adaptive immunity

cGAS-STING signaling pathway from A to Z - Brochures

HTRF Product Catalog

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

Cytokine assays: a guide to success with HTRF

Setting up a successful cytokine assay - Guides

Portfolio of Life Sciences 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

Characterize STING agonists with HTRF human IFNb

Data about HTRF IFNb correlation with gene reporter assay and ELISA - Application Notes

Protocol for Th17 differentiation from CD4 mouse T-lymphocyte

A step by step protocol for Th17 cells differentiation from naive CD4 mouse cells - Technical Notes

An innate and adaptive immunity recap

Insight into the diversity of cells & signaling pathways - Guides

Molecular basis of neuroinflammation and neurodegeneration diseases

The essential guide for extending your knowledge on the molecular mechanisms of neurodegenerative diseases - Guides

Neurodegeneration and its main related diseases

Discover this infographic design on neurodegenerative diseases - Infographics

Neuroinflammation study combining hiPSC-derived astrocytes and HTRF

See published experiments and data demonstrating how immunoassays rise to the challenge of astrocyte studies in neuroinflammation research - Application Notes

On-demand webinar: Linking Neuroinflammation and Neurodegeneration

New insight into neuroinflammation research - Videos

Neutralizing antibodies in Autoimmunity

Power up your autoimmunity research projects with this application note - Application Notes

Assays for autoimmunity research

Advance your research on autoimmune diseases - Flyers

Assays for neurosciences research

Advance your research on neurodegenerative diseases - Flyers

Advance your research on Fibrosis

Kits and reagents for Fibrosis research - Flyers

Manual IL6 (h) Kit / 62hil06peg-62hil06peh

62hil06peg-62hil06peh - Product Insert

Manual IL6 (h) Kit / 62hil06pet

62hil06pet - Product Insert

Certificate of Analysis IL6 (h) Kit / 62HIL06PEG

62HIL06PEG Batch 09E - Quality Control Report

Certificate of Analysis IL6 (h) Kit / 62HIL06PET

62HIL06PET Batch 10B - Quality Control Report

Certificate of Analysis IL6 (h) Kit / 62HIL06PET

62HIL06PET Batch 10C - Quality Control Report

Certificate of Analysis IL6 (h) Kit / 62HIL06PET

62HIL06PET Batch 10D - Quality Control Report

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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