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HTRF Mouse IL-6 Detection Kit, 500 Assay Points

The HTRF mouse IL6 kit is designed for the quantification of mouse 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 mouse IL6 kit is designed for the quantification of mouse 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: 500 Assay Points
Part #:
62MIL06PEG
List Price
USD 1,103.00
Unit Size: 10,000 Assay Points
Part #:
62MIL06PEH
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. When assaying, always follow recommended protocol and avoid highly haemolyzed samples.

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
Mouse
Technology
TR-FRET
Therapeutic Area
Metabolism/Diabetes
NASH/Fibrosis
Neuroscience
Oncology & Inflammation
Unit Size
500 Assay Points

Video gallery

Citations

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.

 

1cytokines-how-it-works-assay-principle-human-il13-62hil13peg-62hil13peh.svg

 

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,
 

Revvity also worked with Myassays.com to help you in your data analysis. You’ll be able to access a free online software to run your IL6 analysis.

 

Assay details

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

 

 

Analytical performance

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.

 

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