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.

Intra-assay (n=24)

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

Inter-assay (n=4)

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.

Anti CD3 and CD28 dose response effect on IL2 secretio

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.