The Mouse IL6 kit is designed for the quantification of mouse IL6 release in cell supernatant.
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  • No-wash
  • Low sample consumption
  • Under 2h bench time
The Mouse IL6 kit is designed for the quantification of mouse IL6 release in cell supernatant.
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In stock

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 the main promoter of T cell differentiation into TH17, a new component of immuno-oncology.

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 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 mouse IL6 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 &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

Intra and inter assay

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.

HTRF Cytokine cell-based assays

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In collaboration with Molecular Devices - Application Notes

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

Validated protocol - Technical Notes

Webinar: NASH - Fibrosis Research and Drug Discovery

Featuring a panel of experts - Videos

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What to expect at the bench - Videos

Product Insert IL6 (m) Kit / 62MIL06PEG-62MIL06PEH

62MIL06PEG-62MIL06PEH - 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

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