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No-Wash Immunoassay for Active IL-18

  • Specific detection of mature/active IL-18; does not recognize pro-IL-18
  • Fast add-and-read protocol
  • Scalable to 96- or 384- well plates, protocols included
  • Pack sizes show assay number for 96-well plates

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Catalog Number: W1910

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Catalog Number: W191A-C

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Overview
Protocols
Specifications
Resources
Related Products

Specific Detection of Active IL-18 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.

Note: The total number of assays per kit will vary based on final well volume.

For standard well volumes:

  • 100 assay kits: applicable for 100 assays in 96-well plates or 400 assays in 384-well plates.
  • 500 assay kits: applicable for 500 assays in 96-well plates or 2,000 assays in 384-well plates.
  • 1,000 assay kits: applicable for 1,000 assays in 96-well plates or 4,000 assays in 384-well plates.

How the Lumit® IL-18 Immunoassay Works

Lumit® Flex technology is a luminescent structural complementation system composed of two small peptide tags (Peptide α and Peptide β) and a reagent-based polypeptide (Lumit® Flex Detection Protein). In the Lumit® Active IL-18 Immunoassay, a monoclonal anti–human IL-18 antibody and human IL-18 binding protein (IL-18BP) are labeled with Peptide α and Peptide β, respectively. In the presence of active IL-18, the two peptides are brought into proximity. When Lumit® Detection Reagent B (containing the Flex Detection Protein) is added, a functional luciferase is reconstituted and produces a bright luminescent signal in the presence of Lumit® Substrate. The signal is directly proportional to active IL-18 levels.

The assay specifically detects active IL-18 and does not recognize pro-IL-18 or IL-18 bound to endogenous IL-18BP.

For proper assay performance, hIL-18BP-β must be incubated with cells during treatment to capture active IL-18 immediately as it is released from cells. This is because active IL-18 is unstable in cell culture media. To see data demonstrating the instability of active IL-18, review this scientific poster.

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

Lumit® IL-18 Immunoassay protocol schematic.

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

Dynamic Range

7.8pg/ml–5,714pg/ml

Limit of Detection (LOD)

1.9pg/ml (3 SD above background)

Minimal Detectable Dose (MDD)

1.6pg/ml (2 SD above background)

Assay Time

70 minutes

Sample Type

Cell culture supernatants

SD=Standard Deviation

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

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.

Specifications

Catalog Number:

Contenido

Item Part # Presentación

Lumit® Detection Substrate B

VB405A 1 × 160μl

Lumit® Detection Buffer B

VB406A 1 × 3.2ml

Lumit® Flex Detection Protein

VB720A 1 × 450μl

hIL-18BP-Peptide β, 1000X

W192A 1 × 20μl

Anti-hIL-18 mAb-Peptide α, 1000X

W193A 1 × 20μl

Lumit® Human Active IL-18 Standard

W191A-C 1 × 25μl

Certificado de Análisis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Condiciones de Almacenaje

AA

Patentes y Exclusiones

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 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.

Contenido

Item Part # Presentación

Lumit® Detection Substrate B

VB405B 1 × 1.25ml

Lumit® Detection Buffer B

VB406B 1 × 25ml

Lumit® Flex Detection Protein

VB720B 1 × 4.5ml

hIL-18BP-Peptide β, 1000X

W192B 1 × 200μl

Anti-hIL-18 mAb-Peptide α, 1000X

W193B 1 × 200μl

Lumit® Human Active IL-18 Standard

W191A-C 1 × 25μl

Certificado de Análisis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Condiciones de Almacenaje

AA

Patentes y Exclusiones

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 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.

Contenido

Item Part # Presentación

Lumit® Detection Substrate B

VB405A 5 × 160μl

Lumit® Detection Buffer B

VB406A 5 × 3.2ml

Lumit® Flex Detection Protein

VB720A 5 × 450μl

hIL-18BP-Peptide β, 1000X

W192A 5 × 20μl

Anti-hIL-18 mAb-Peptide α, 1000X

W193A 5 × 20μl

Lumit® Human Active IL-18 Standard

W191A-C 1 × 25μl

Certificado de Análisis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Condiciones de Almacenaje

AA

Patentes y Exclusiones

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 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.

Contenido

Item Part # Presentación

Lumit® Human Active IL-18 Standard

W191A-C 1 × 25μl

Certificado de Análisis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Condiciones de Almacenaje

AA