Human IL5 Assay Kit

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.
Cisbio's Human IL5 kit is a homogeneous, add and read assay to measure IL5 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) 15 pg/mL & 71 pg/mL
Range 71 – 10,000 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
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 IL5 analysis.

 

How to use HTRF cytokine assays

How to measure multiple cytokines from 1 sample

Analytical performance

Precision

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.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Human IL5 kit - 500 tests62HIL05PEG
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Human IL5 kit - 10,000 tests62HIL05PEH
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Companion products

DescriptionCat. noProduct insertMSDS
Human IL5 kit - 3 Standards62HIL05CD3
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Human IL5 kit - Standard62HIL05CDA
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Documents