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 Revvity 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.

Human IL-1β standard curve

Intra-assay (n=24)

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

Inter-assay (n=4)

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

Modulation of secretion of human IL-1β in THP1 cell line

THP1 cells plated at 25, 50, and 100 kcells/well were stimulated for 18h with PMA and LPS, respectively used at 50 ng/mL and 1 µg/mL. 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed with the Human IL-1ß Assay.

IL-1ß secretion in PBMC stimulated with LPS

PBMC plated at 50, 100, and 200 kcells/well were stimulated for 18 h with increasing concentrations of LPS ranging from 0.02 to 2 µg/mL. Then 16 µL of supernatants were transferred into a white detection plate (384 low volume) to be analyzed using the Human IL-1ß assay kit.

Effect of dexamethasone on PBMC induced IL-1ß release

PBMC plated at 50 and 100 kcells/well were stimulated for 18h with 0.2 µg/mL of LPS and a co-treatment of increasing concentrations of dexamethasone, ranging from 0.05 to 50 µg/mL. Then 16 µL of supernatants were transferred into a white detection plate (384 low volume) to be analyzed using the Human IL-1ß Assay Kit.