Human IL2 Assay Kit

IL2's major role is to modulate cell-mediated immunity through its effect on T cells. IL2 promotes the differentiation of T cells into effector or memory T cells, thus helping the body fight off infection.
IL2 is also a key player in immuno-oncology since it is the main promoter of the differentiation of T cells into Treg.
Cisbio's Human IL2 kit is a homogeneous, add and read assay to measure IL2 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) 7 pg/mL & 37 pg/mL
Range 37 – 8,000 pg/mL
Time to result 3h at RT
Calibration NIBSC (86/500) value (IU/mL) = 0,01 x HTRF hIL2 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 IL2 analysis.

 

How to use HTRF cytokine assays

How to measure multiple cytokines from 1 sample

Analytical performance

Precision

Intra-assay (n=24)

Sample Mean [IL2] (pg/mL) CV
1 75 7%
2 2997 3%
3 6136 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 [IL2] (pg/mL) Mean (delta R) CV
1 98 346 12%
2 432 1576 12%
3 1904 5911 10%
    Mean CV 11%

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

Assay validation

1. IL2 secretion in Jurkat cells stimulated with anti CD3&CD28: cell densities

Jurkat cells plated @ 50 and 100 kcells/well were stimulated for 24 h with anti CD3 & and anti CD28 respectively prepared at 10 & 1 µg/mL.
16 µL of supernatants were then transferred into a white detection plate to be analyzed by the Human IL2 Assay Kit.

2. IL2 secretion in Jurkat cells stimulated with anti CD3&CD28: agonist dose response

Jurkat cells plated @ 200 kcells/well were stimulated for 24 h with anti CD3 and anti-CD28 used at various concentrations:
0, 0.5, 1, 10 µg/mL for the anti CD3 and 0, 1, 5, 10, 20 µg/mL for the anti CD28.
16 µL of supernatants were then transferred into a white detection plate to be analyzed by the Human IL2 Assay Kit.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Human IL2 kit - 500 tests62HIL02PEG
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Human IL2 kit - 10,000 tests62HIL02PEH
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Human IL2 kit - 1 x 96 tests62HIL02PET
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Companion products

DescriptionCat. noProduct insertMSDS
Human IL2 kit - 3 Standards62HIL02CD3
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Human IL2 kit - Standard62HIL02CDA
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