Lumit® HMGB1 Human/Mouse Immunoassay
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
Catalog Number:
Choose a product
Size
Catalog Number: W6110
Catalog Number: W6112
Catalog Number: W140A-C
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.
|
Target |
Human HMGB1 |
Mouse HMGB1 |
|
Limit of Detection |
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
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
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
Specifications
Catalog Number:
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 SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
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.
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 SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
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.
What's in the box?
| Item | Part # | Size |
|---|---|---|
HMGB1 (Human) Positive Control |
W140A | 1 × 20μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
Resources
Articles
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Frequently Used With
RealTime-Gloâ„¢ Extracellular ATP Assay
Bioluminescent assay designed for continuous, kinetic monitoring of ATP released from dying, stressed or activated cells.
GA5010, GA5011, GA5012
CellToxâ„¢ Green Cytotoxicity Assay
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Antibody labeling kit and detection reagents to help develop Lumit® Immunoassays.
VB2500, VB2010, VB2020, VB2030, VB4050, VB4060
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High-performance microplate reader for detecting luminescence, fluorescence and absorbance.
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