HTRF Human IFN-β Detection Kit, 500 Assay Points

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.

Intra assay (n=24)

Inter assay (n=4)

STING pathway activation by cGAMP triggers hIFNβ release in PBMC

400,000 PBMC were seeded in a 96 well plate under 100 µL of cell culture medium, then stimulated with 50 µL of cGAMP used at 67µM. After an overnight incubation, 14 µL of supernatant were transferred into a white detection plate and human IFNβ was quantified by HTRF.

​LPS efficiently induces hIFNβ release in THP1 cells

400,000 THP1cells were seeded in a 96 well plate under 100 µL, then stimulated with 50 µL of LPS used at 20 µg/ml. After 4h30 incubation, 14 µL of supernatant were transferred into a white detection plate, and human IFNβ was quantified by HTRF.

Recombinant THP1 cells expressing constitutively activated STING variant spontaneously releases hIFNβ

400,000 of THP1-WT cells (wild type) and THP1-S154 cells (THP1-Dual™ KI-hSTING-S154 Cells, Invivogen) were seeded in a 96 well plate, under 100 µL of cell culture medium. After 4 h 30 of stimulation, 14 µl of supernatant were transferred into a white detection plate, and human IFNβ was quantified by HTRF.

dsDNA mimetic (CDS agonist) induces hIFNβ secretion

400,000 THP1-S154 cells (THP1-Dual™ KI-hSTING-S154 Cells, Invivogen) were seeded in a 96 well plate under 100 µL of cell culture medium, then treated with dsDNA mimetic (CDS Agonist – LyoVec, Invivogen). After an overnight incubation, 14 µL of supernatant were transferred into a white detection plate, and human IFNβ was quantified by HTRF.