Mouse IL6 Assay Kit

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 has been studied in a wide variety of research areas including Diabetes, Alzheimer’s, depression and several cancers.
Along with TGFb, IL6 is the main promoter of T cell differentiation into TH17, a new component in immuno-oncology.
Cisbio's Mouse IL6 kit is a homogeneous, add and read assay to measure IL6 secretion in supernatant. It offers a fast, efficient alternative to ELISA.

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 & 19 pg/mL
Range 19 – 7,000 pg/mL
Time to result 1h at RT
Calibration NIBSC (93/730) value (IU/mL) = 0,7 x HTRF mIL6 value (pg/mL)
Species Mouse only
Download Insider tips for PBMC isolation Technical Note
Download cytokine release from fresh blood samples application notes

Assay principle

The assay protocol, using a Cisbio 96-well low volume or a 384-well small volume white plate (20 µL final), is described below. Cell supernatant, sample or standard are dispensed directly into the assay plate for the detection by HTRF® reagents. 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 in 96 to 1536-well plates by simply resizing each addition volume proportionally.

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.

Add

Cytokines assay principle add

Read

Cytokines assay principle read

(1) Assay data analysis:

  • "Guidelines for data reduction" button will show you how dealing with HTRF data processing and 4PL 1/y2 fitting.
  • "MyAssays" button will direct you to a free online software able to run IL6 analysis.

 

How to use HTRF cytokine assays

How to measure multiple cytokines from 1 sample

Analytical performance

Precision

Intra-assay (n=24)

Sample Mean [IL6] (pg/mL) CV
1 40 12%
2 1400 7%
3 2800 11%
  Mean CV 10%

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 72 503 6%
2 448 3188 10%
3 2800 18091 10%
    Mean CV 9%

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

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Mouse IL6 kit - 500 tests62MIL06PEG
Mouse IL6 kit - 10,000 tests62MIL06PEH
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Companion products

DescriptionCat. noProduct insertMSDS
Mouse IL6 kit - 3 Standards62MIL06CD3
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Mouse IL6 kit - Standard62MIL06CDA
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