Fast add-and-read protocol performed directly on cells
Luminescent detection with just a plate-reading luminometer
Scalable to 96- or 384-well plates
Quantify Cytokine Release Without Tedious Wash Steps
The Lumit™ IL-6 Immunoassay quantitatively measures released IL-6 in cell culture samples using a simple, no-wash protocol. Just add labeled antibodies to the sample, add detection reagent and read luminescent signal using a standard plate-reading luminometer. The entire protocol is completed in less than 70 minutes! The assay can be used directly on cells in culture or culture medium transferred to a separate assay plate.
How the Lumit™ IL-6 Immunoassay Works
Primary antibodies to IL-6 were selected for their specific and sensitive detection and labeled with the LgBiT and SmBiT subunits of NanoBiT® Luciferase. In the presence of IL-6, the subunits are brought together to form an active luciferase enzyme. Addition of optimized substrate generates a bright luminescent signal proportional to IL-6 levels.
Simple One-Plate Protocol, No Wash Steps
Large Dynamic Range of Lumit™ IL-6 Detection
The pg/ml to ng/ml linear range of Lumit™ detection covers expected concentrations for most cell models.
Assay
IL-6 Human Immunoassay
Limit of Detection (LOD)
7.5pg/ml (3 SD above background)
Dynamic Range
18.2pg/ml – 25ng/ml
Minimal Detectable Dose (MDD)
5.0pg/ml (2 SD above background)
Assay Time
70 minutes
Sample Type
Cell culture supernatants
SD=Standard Deviation
Rapid Detection of IL-6 Release from PBMCs
The Lumit™ IL-6 (Human) Immunoassay (left) and standard ELISA (right) were used to quantify IL-6 release from PBMCs treated with three inducers (cell stimulation cocktail, lipopolysaccharide or R848) for up to 72 hours. The results are similar, while the Lumit™ Immunoassay was completed in one fifth the time of the ELISA, with no transfer and wash steps or plate washer needed.
