Lumit® HMGB1 Human/Mouse Immunoassay

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No-Wash Immunoassay for HMGB1 Detection

  • Fast add-and-read assay performed directly on cells
  • Detects human or mouse HMGB1
  • Luminescent detection with just a plate-reading luminometer

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Catalog number selected: W6110

$ 597.00
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This product is available through the Promega Helix onsite stocking program. We offer numerous convenient solutions to meet your lab's needs.

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Lumit® HMGB1 Human/Mouse Immunoassay
100 assays
$ 597.00
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Fast, No-Wash Detection of HMGB1

High-Mobility Group Box 1 (HMGB1) functions as a damage-associated molecular pattern (DAMP) released from cells in inflammatory responses. HMGB1 is often measured as a key biomarker of immunogenic cell death, a specialized form of cell death that results in an immune response. HMGB1 also functions as a proinflammatory cytokine. Current methods to detect HMGB1, such as ELISA, require tedious wash steps and lengthy protocols. Lumit® technology simplifies HMGB1 quantitation with an add-mix-measure protocol completed in 90 minutes or less.

How the Lumit® HMGB1 Immunoassay Works

Primary antibodies to HMGB1, selected for their specific and sensitive detection, are labeled with the LgBiT and SmBiT subunits of NanoBiT® Luciferase. In the presence of HMGB1, the subunits are brought together to form an active luciferase enzyme. Addition of optimized substrate generates a bright luminescence signal proportional to HMGB1 levels. The assay detects both human and mouse HMGB1.

How the lumit HMGB1 assay works.

Simple One-Plate Protocol, No Wash Steps

Lumit® Immunoassay Workflow

Large Dynamic Range of HMGB1 Detection

Due to the homology of human and mouse HMGB1, the kit can be used for both species with minimal performance differences.

Lumit™ HMGB1 Human Immunoassay standard curve.
Lumit™ HMGB1 Mouse Immunoassay standard curve.

Target

Human HMGB1

Mouse HMGB1

Limit of Detection 
(3SD above background)

1ng/ml 

3ng/ml 

Dynamic Range

4–1,000ng/ml

3–2,187ng/ml

Assay Time

90 minutes

Sample Type

Cell culture samples

SD=Standard Deviation

Dual DAMP Measurements in a Human Cell Line

Data showing ATP release and HMGB1 release after mitoxantrone treatment

In parallel assay plates, U937 cells were dosed with mitoxantrone and assayed for extracellular ATP release (left) kinetically over 24 hours using the RealTime-Glo™ Extracellular ATP Assay. Lumit® HMGB1 Immunoassay reagents were added directly to the second plate of treated cells after 24 hours of treatment. Mitoxantrone produces a dose- and time-dependent release of extracellular ATP (left) and a dose-dependent release of HMGB1 (right) with potency agreement between the two measurements of damage-associated molecular patterns.

Detect Drug-Induced Immunogenic Cell Death in Various Cell Culture Models

Drug-induced immunogenic cell death detected with HMGB1 assay.

Human U2OS cells (left) and mouse EL4 cells (right) were treated with doxorubicin for 24 hours. After 24 hours, the Lumit® HMGB1 Immunoassay was performed directly on cell culture samples to measure HMGB1 released from treated cells.

Also Available: Live-Cell Kinetic Measurement of Extracellular ATP

The RealTime-Glo™ Extracellular ATP Assay kinetically measures the release of ATP from dying cells so you can assess immunogenic cell death induction in real time. Use this assay to complement the Lumit® HMGB1 Immunoassay for a more complete picture of immunogenic cell death.

Protocols

Complete Protocol

Specifications

You are viewing: W6110 Change Configuration

What's in the box?

Item Part # Size

HMGB1 (Human) Positive Control

 W140A 1 × 20μl

Anti-hHMGB1 mAb-SmBiT, 250X

 W141A 1 × 50μl

Anti-hHMGB1 mAb-LgBiT, 250X

 W142A 1 × 50μl

Lumit® Detection Substrate B

 VB405A 1 × 160μl

Lumit® Detection Buffer B

 VB406A 1 × 3.2ml

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

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

Storage Conditions

BB

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.

Specifications

You are viewing: W6112 Change Configuration

What's in the box?

Item Part # Size

HMGB1 (Human) Positive Control

 W140A 1 × 20μl

Anti-hHMGB1 mAb-SmBiT, 250X

 W141A 5 × 50μl

Anti-hHMGB1 mAb-LgBiT, 250X

 W142A 5 × 50μl

Lumit® Detection Substrate B

 VB405A 5 × 160μl

Lumit® Detection Buffer B

 VB406A 5 × 3.2ml

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

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

Storage Conditions

BB

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

Posters

Other Resources

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