Human IL8 kit HTRF®

The HTRF human IL8 kit is designed for the quantification of human IL8 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 IL8 kit is designed for the quantification of human IL8 release in cell supernatant.



Also called CXCL8, IL8 is a chemokine mainly produced by macrophages, T cells, and neutrophils. IL8 acts as a chemoattractant for neutrophils, and promotes infiltration and activation at inflammation sites. IL8 is involved in several cancer types due to its ability to promote angiogenesis and cell proliferation, as well as to inhibit apoptosis.

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-8 AlphaLISA 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.
IL8 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 IL8 analysis.

Technical specifications of human IL8 kit

Sample size 16 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, buffers &protocol.
LOD &LOQ (in Diluent) 6 pg/mL &32 pg/mL
Range 32 – 4,000 pg/mL
Time to result 1h at RT
Calibration NIBSC (89/520) value (IU/mL) = 0,01 x HTRF hIL8 value (pg/mL)
Species Human only

Intra and inter assay

Intra-assay (n=24)

Sample Mean [IL8] (pg/mL) CV
1 58 9%
2 314 4%
3 2074 2%
Mean CV 5%

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

Inter-assay (n=4)

Sample [IL8] (pg/mL) Mean (delta R) CV
1 78 273 2%
2 376 1504 12%
3 1818 5990 5%
Mean CV 6.3%

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

Dilutional linearity

Dilution factor[IL8] expected (pg/mL)[IL8] detected (pg/mL)Recovery

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

Spike and recovery

[IL8] added (pg/mL)[IL8] expected (pg/mL)[IL8] detected (pg/mL)Recovery

​The same amount of recombinant cytokine was added to 2 different serum samples, and the set of responses obtained from a standard curve was compared to the calculated expected values. The ~ 100% of recovery found validates the sample matrix used for this assay.

Inhibition of IL8 secretion in THP1 cells with JTE607

THP1 cells plated at 100 kcells/well were incubated with increasing concentrations of JTE 607 for 18h, then stimulated for 3 h with 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 IL8 Assay.

Inhibition of IL8 secretion in THP1 cells with JTE607.

IL8 secretion in PBMCs stimulated with LPS

PBMC plated at 50, 100, 200, and 400 kcells/well were stimulated for 3 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 IL8 Assay.

IL8 secretion in PBMCs stimulated with LPS.

Celular High Throughput Screening assays using HTRF detection of cytokines

In collaboration with GSK - Scientific Presentations

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

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

How to measure multiple cytokines from 1 sample

What to expect at the bench - Videos

Product Insert IL8 (h) Kit / 62HIL08PEG-62HIL08PEH

62HIL08PEG-62HIL08PEH - Product Insert

Product Insert IL8 (h) Kit / 62HIL08PET

62HIL08PET - Product Insert

HTRF makes cytokine detection easy

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

HTRF Product Catalog

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

Cytokine Portfolio Flyer

Discover no-wash cytokine assay solutions - Flyers

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

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

Product Insert IL8 (h) Kit / 62HIL08PET-v6

62HIL08PET-v6 - Product Insert

Cytokine assays: a guide to success with HTRF

Setting up a successful cytokine assay - Guides

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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