Lumit® Active IL-18 (Human) Immunoassay

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No-Wash Immunoassay for Active IL-18 Detection

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Lumit® Active IL-18 (Human) Immunoassay

Quantify Inflammasome Activation Without Tedious Wash Steps

The Lumit® Active IL-18 Immunoassay quantitatively measures active IL-18 in cell culture samples using a simple, no-wash protocol. Just add labeled binding protein with treatment, add labeled antibody, 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 samples transferred to a separate assay plate.

How the Lumit® IL-18 Immunoassay Works

The Lumit® IL-18 Immunoassay uses one primary antibody to IL-18, selected for its specificity and sensitivity, and labeled with the LgBiT subunit of NanoBiT® luciferase in combination with the IL-18 Binding Protein, labeled with the SmBiT subunit of NanoBiT® Luciferase. In the presence of cleaved, mature IL-18, the NanoBiT® subunits are brought together to form an active luciferase enzyme. Addition of the optimized luciferase substrate generates a bright luminescent signal proportional to active IL-18 levels. The assay is specific to active IL-18 and does not detect pro-IL-18 or inactive IL-18 bound by endogenous IL-18 binding protein.

Schematic of Lumit™ IL-18 Immunoassay concept showing anti-IL-18-LgBiT and SmBiT-IL-18 binding protein binding to IL-18 and forming NanoBiT® Luciferase.

Simple One-Plate Protocol Requires No Wash Steps

assay protocol part 1
assay protocol part 2

Broad Linear Range of Lumit® IL-18 Detection

The picomolar to nanomolar linear detection range of Lumit® IL-18 Immunoassay covers expected concentrations for most cell models.

Graph showing standard curve for Lumit™ IL-18 (Human) Immunoassay.

Specification

Lumit® IL-18 Immunoassay

Limit of Detection

≤10pg (3 SD above background)

Dynamic Range

11pg–20ng

Assay Time

70 minutes

Sample Type

Cell culture supernatants

SD=Standard Deviation

Featured Webinar

Using Lumit® Technology to Address Inflammasome-Mediated Cytokine Release

In this webinar, Promega Sr. Research Scientist, Dr. Martha O’Brien, describes developing tools to measure inflammasome activation, including the Lumit® Active IL-18 Immunoassay and Lumit® IL-1β Immunoassay. Dr. O’Brien shows how Lumit® Assays can be multiplexed with other inflammasome assays to identify and rank the potency of inflammasome inhibitors.

View On-Demand Webinar

Want to see all the data?

Download the Scientific Poster

Detect IL-18 Release Following Inflammasome Activation

Bar graph showing active IL-18 detection in cells treated with Nigericin and LPS + Nigericin.

The Lumit® Active IL-18 (Human) Immunoassay was used to quantitate secretion of active IL-18 from THP-1 cells following treatment. Active IL-18 is found in cell culture media samples following treatment with either nigericin or nigericin + LPS but not in untreated cells or cells treated with LPS alone. The amount of active IL-18 secreted was calculated using the standard curve. Data shown is an average of 3 replicates; error bars represent standard deviation.

Looking for additional tools to monitor inflammasome activation? Visit our Inflammasome Activation Page.

Patents and Disclaimers

U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.

U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 10,648,971 and other patents and patents pending.

Resources

Articles

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