The HTRF human IL5 kit is designed for the quantification of human IL5 release in cell supernatant.

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  • No-wash No-wash
  • Low sample consumption Low sample consumption
  • Under 2h bench time Under 2h bench time

The HTRF human IL5 kit is designed for the quantification of human IL5 release in cell supernatant.

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Overview

IL5 is mainly produced by TH2 cells upon IL4 activation, but also by eosinophils and mast cells. IL5 promotes B cell growth and stimulates immunoglobulin secretion. It also induces eosinophil activation, proliferation, and differentiation.


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-5 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.

Benefits

  • SCALABLE FROM 96 TO 1536WELLS
  • TH2 CELLS ACTIVATION READOUT

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.
IL5 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 IL5 analysis.

Technical specifications of human IL5 kit

Sample size 16 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, buffers &protocol.
LOD &LOQ (in Diluent) 15 pg/mL &71 pg/mL
Range 71 – 20,000 pg/mL
Time to result 2h at RT
Calibration NIBSC (90/586) value (IU/mL) = 0,012 x HTRF hIL5 value (pg/mL)
Species Human only

Intra and inter assay

Intra-assay (n=24)

Sample Mean [IL5] (pg/mL) CV
1 104 9%
2 969 2%
3 8064 2%
Mean CV 4%

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

Inter-assay (n=4)

Sample [IL5] (pg/mL) Mean (delta R) CV
1 388 552 4%
2 1878 2413 4%
3 4131 5060 4%
Mean CV 4%

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

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Product Insert IL5 (h) Kit / 62HIL05PEG-62HIL05PEH

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Cisbio Product Catalog 2019

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A guide to Homogeneous Time Resolved Fluorescence

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