Lumit® HMGB1 Human/Mouse Immunoassay

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.

Simple One-Plate Protocol, No Wash Steps

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.

Protocolos

Complete Protocol

Lumit™ HMGB1 (Human/Mouse) Immunoassay Technical ManualPDF (1981 KB) – English

Contenido

Item	Part #	Presentación
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

Certificado de Análisis

Search by lot number

Use Restrictions

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

Condiciones de Almacenaje

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
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

Certificado de Análisis

Search by lot number

Use Restrictions

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

Condiciones de Almacenaje

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.

Recursos

Artículos

Posters

Improved DAMP Assays for the In Vitro Assessment of Immunogenic Cell Death

Otros

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Lumit® HMGB1 Human/Mouse Immunoassay

No-Wash Immunoassay for HMGB1 Detection

Fast, No-Wash Detection of HMGB1

How the Lumit® HMGB1 Immunoassay Works

Simple One-Plate Protocol, No Wash Steps

Large Dynamic Range of HMGB1 Detection

Dual DAMP Measurements in a Human Cell Line

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

Also Available: Live-Cell Kinetic Measurement of Extracellular ATP

Protocolos

Complete Protocol

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Recursos

Artículos

Posters

Otros

Lumit® HMGB1 Human/Mouse Immunoassay

No-Wash Immunoassay for HMGB1 Detection

Fast, No-Wash Detection of HMGB1

How the Lumit® HMGB1 Immunoassay Works

Simple One-Plate Protocol, No Wash Steps

Large Dynamic Range of HMGB1 Detection

Dual DAMP Measurements in a Human Cell Line

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

Also Available: Live-Cell Kinetic Measurement of Extracellular ATP

Protocolos

Complete Protocol

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Recursos

Artículos

Posters

Otros

Productos Similares

Lumit® IL-1β Human/Mouse Immunoassay

Lumit® Immunoassay Labeling Kit and Detection Reagents

Lumit® IL-6 (Human) Immunoassay

Lumit® TNF-α (Human) Immunoassay

Usado con frecuencia junto con

RealTime-Glo™ Extracellular ATP Assay

GloMax® Discover System

CellTox™ Green Cytotoxicity Assay

Caspase-Glo® 1 Inflammasome Assay

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