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No-Wash Immunoassay for IL-17A Detection​

  • ​Fast add-and-read protocol performed directly on cells​
  • Luminescent detection with any microplate luminometer​
  • Scalable to 96- or 384-well plates​

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Overview
Protocols
Specifications
Resources
Related Products

Quantify IL-17A Release Without Tedious Wash Steps

IL-17A, a pro-inflammatory cytokine, is a pivotal player in inflammation as it mediates the host’s defense against pathogens and is involved in the pathogenesis of various inflammatory and autoimmune diseases. Therapeutically targeting IL-17A shows a reduction in inflammation and improves clinical outcomes in autoimmune conditions.

The Lumit® IL-17A Immunoassay quantitatively measures released IL-17A 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-17A Immunoassay Works

Primary antibodies to IL-17A were selected for their specific and sensitive detection and labeled with the LgBiT and SmBiT subunits of NanoBiT® Luciferase. In the presence of IL-17A, the subunits are brought together to form an active luciferase enzyme. Addition of optimized substrate generates a bright luminescent signal proportional to IL-17A levels. The assay is specific to IL-17A and does not detect B, D, E and F family members.

Schematic of Lumit™ IL-17A Immunoassay concept showing LgBiT and SmBiT binding protein binding to IL-17 and forming NanoBiT® Luciferase.

Simple One-Plate Protocol Requires No Wash Steps

Lumit™ IL-17A (Human) Immunoassay workflow.

Broad Linear Range of Lumit® IL-17A Detection

The picomolar to nanomolar linear detecton range of Lumit® IL-17A Immunoassay covers expected concentrations for most cell models.

Graph showing standard curve for Lumit® IL-17A (Human) Immunoassay.

Specification

Lumit® IL-17A Immunoassay

Dynamic Range

18.2–25,000pg/ml

Limit of Detection (LOD)

2.2pg/ml (3 SD above background)

Minimal Detectable Dose (MDD)

1.3pg/ml (2 SD above background)

Assay Time

70 minutes

Sample Type

Cell culture supernatants

SD=Standard Deviation

 

Need to measure a different target?

View our growing portfolio of
Lumit® Immunoassays.

For a biotech that understands time is of significant value, the Lumit® Immunoassays are an essential way for us to gather reliable data fast so that we can reach conclusions and make decisions sooner.

Jonathan Chow, Principal Scientist, Corner Therapeutics

Optimizing the Lumit® IL-6 Immunoassay for High-Throughput Drug Screening

In this webinar, we share key considerations for scaling Lumit® Immunoassays from a single 96-well plate to many 364-well plates and offer tips for optimizing Lumit® Immunoassays for HTS applications. You will also hear Galapagos scientist Dr. Nick Verstraeten share how his team used the Lumit® IL-6 Immunoassay in a screening campaign for antiviral drugs.

View On-Demand Webinar

Lumit® Detection of IL-17A Release from PBMCs

Line graph showing Lumit® detection of IL-17A release from PBMC.

 

Peripheral Blood Mononuclear Cells (PBMC) were plated in 96-well plate format at 100,000 cells/well in RPMI 1640 + 10% heat-inactivated FBS. Cells were treated in quadruplicate for 48 hours with increasing concentrations of cell stimulation cocktail (a mixture of PMA and ionomycin). Following treatment, IL-17A release was quantitated using the Lumit® IL-17A Immunoassay.

Specifications

Catalog Number:

Contenido

Item Part # Presentación

Lumit® Detection Substrate B

VB405A 1 × 160μl

Lumit® Detection Buffer B

VB406A 1 × 3.2ml

Anti-hIL-17A mAb-SmBiT, 500X

W144A 1 × 30μl

Anti-hIL-17A mAb-LgBiT, 500X

W145A 1 × 30μl

Human IL-17A Standard

W143A 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 10,648,971 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

Anti-hIL-17A mAb-SmBiT, 500X

W144B 1 × 300μl

Anti-hIL-17A mAb-LgBiT, 500X

W145B 1 × 300μl

Human IL-17A Standard

W143A 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 10,648,971 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

Anti-hIL-17A mAb-SmBiT, 500X

W144A 5 × 30μl

Anti-hIL-17A mAb-LgBiT, 500X

W145A 5 × 30μl

Human IL-17A Standard

W143A 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 10,648,971 and other patents and patents pending.

Contenido

Item Part # Presentación

Human IL-17A Standard

W143A 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