The HTRF human IL2 kit is designed for the quantification of human IL2 release in cell supernatant.

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  • No-wash No-wash
  • Validated on PBMC Validated on PBMC
  • Under 2h bench time Under 2h bench time

The HTRF human IL2 kit is designed for the quantification of human IL2 release in cell supernatant.

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Overview

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 (regulatory T cells).


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-2 AlphaLISA kitand the  Human IL-2 AlphaLISA biotin-free 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

  • VALIDATED ON PBMC
  • T CELL 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.
IL2 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 IL2 analysis.

Technical specifications of human IL2 kit

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

Intra and inter assay

Intra-assay (n=24)

SampleMean [IL2] (pg/mL)CV
1757%
229973%
361362%

Mean CV4%

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.

IL2 secretion in Jurkat cells stimulated with anti CD3 and CD28

Jurkat cells plated at 50 and 100 kcells/well were stimulated for 24 h with anti CD3 & and anti CD28 respectively prepared at 10 and 1 µg/mL. 16 µL of supernatants were transferred into a white detection plate (384 low volume) to be analyzed by the Human IL2 Assay.

IL2 secretion in Jurkat cells stimulated with anti CD3 and CD28.

Anti CD3 and CD28 dose response effect on IL2 secretion

Jurkat cells plated at 200 kcells/well were stimulated for 24 h with increasing concentrations of anti CD3 and anti-CD28: 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 (384 low volume) to be analyzed by the Human IL2 Assay.
Anti CD3 and CD28 dose response effect on IL2 secretion.
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