Skip to main content
Menu
US

HTRF Human IL-6 Detection Kit, 500 Assay Points

The HTRF human IL6 kit is designed for the quantification of IL6 release in 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 and disposal requirements under European REACH regulations (EC 1907/2006).

Feature Specification
Application Protein Quantification
Sample Volume 16 µL

The HTRF human IL6 kit is designed for the quantification of IL6 release in 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 and disposal requirements under European REACH regulations (EC 1907/2006).

Product Variants
Unit Size: 96 Assay Points
Part #:
62HIL06PET
List Price
USD 652.48
Unit Size: 500 Assay Points
Part #:
62HIL06PEG
List Price
USD 1,103.00
Unit Size: 10,000 Assay Points
Part #:
62HIL06PEH
List Price
USD 10,290.00

Overview

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 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 detection of low levels of analytes in serum or plasma.

Specifications

Application
Protein Quantification
Brand
HTRF
Detection Modality
HTRF
Product Group
Kit
Sample Volume
16 µL
Shipping Conditions
Shipped in Dry Ice
Target Class
Cytokines
Target Species
Human
Technology
TR-FRET
Therapeutic Area
Metabolism/Diabetes
NASH/Fibrosis
Neuroscience
Oncology & Inflammation
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.

Assay principle human il6
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.
 

Assay details

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

 

Analytical performance

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
Sample Dilution factor [IL6] expected (pg/mL) [IL6] detected (pg/mL) Recovery
1 1 - 2015 -
4.3 469 489 104%
7.4 271 292 108%
10.7 188 210 111%
Mean 108%
2 1 - 354 -
4.3 83 82 101%
7.4 45 48 94%
Mean 97%


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

Spike and recovery
Sample [IL6] added (pg/mL) [IL6] expected (pg/mL) [IL6] detected (pg/mL) Recovery
1 3250 4098 4103 100%


​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.

Assay validation

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.

assay validation human il6
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.

assay validation human il6

 

Resources

Are you looking for resources, click on the resource type to explore further.

1-1 of 1 Resources
Guide Icon
Guide
HTRF solutions, guide to major applications

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

Scroll Icon