D. Steady-Glo ® Luciferase Assay Buffer E. Steady-Glo ® Luciferase Assay Substrate (contains DTT)

Intended Use: The Kit is intended for use as an in vitro test for the quantitative determination of Human Interferon Alpha (IFN-α) bioactivity (IU/ml) using luciferase generated-bioluminescence. Though this kit is provided with Human IFN Alpha 2b as the standard, the kit is also useful for examining the bioactivity of other interferons that bind to and activate the Type I receptor. These include the variety of IFN-α isoforms, IFN-β, IFN-κ and IFN-ω. This test is for research use only. Background: Type I Interferons constitute a family of cytokines comprising at least 18 IFN-α isoforms, and single IFN-β, IFNκ and IFN-ω isotypes identified by their ability to protect cells against viral infection. They are produced by a wide variety of cells in response to a variety of stimuli, most notably in response to viral infection via the interaction of single-stranded or double-stranded viral RNA with Toll-like receptors (TLR) TLR8 and TLR3, respectively1. Assay Principle: The iLiteTM Human IFN alpha assay is a quantitative Gene Reporter Bioassay. The test procedure is based on the sequential addition of diluent and sample to Human Type I interferon-sensitive cells in a bioluminescence microtiter plate. After incubation and addition of lysis reagent/substrate, the resultant bioluminescence intensity is proportional to the amount of Type I interferon activity (IU/ml) in the sample. The assay range is 1.5 IU/ml to 25 IU/ml or 6-200 IU/ml depending on the incubation period chosen