THP1-HMGB1-Lucia™ Cells
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Cat.code:
thp-gb1lc
- Documents
ABOUT
HMGB1-Lucia reporter assay for pyroptosis and necroptosis monitoring
High mobility group box 1 (HMGB1) plays a critical role in the stress response not only inside the cell as a DNA chaperone and cell death regulator but also outside the cell as a prototypical alarmin. Indeed, during non-apoptotic programmed or regulated cell death, such as pyroptosis and necroptosis, HMGB1 is released along with other pro-inflammatory molecules [1-3].
More details about Pyroptosis and necroptosis
THP1-HMGB1-Lucia™ cells are designed to monitor necroptosis and inflammasome-mediated pyroptosis, two forms of necrotic cell death characterized by the release of the alarmin HMGB1 upon cell membrane rupture [1-3]. These cells are derived from the THP-1 human monocytic cell line and stably express a HMGB1::Lucia-luciferase fusion protein. They respond to commonly used inflammasome inducers and necroptosis cocktail inducers. The release of the HMGB1::Lucia protein in the extracellular milieu upon pyroptosis or necroptosis can be readily measured using QUANTI-Luc™ 4 Lucia/Gaussia, a Lucia and Gaussia luciferase detection reagent.
Key features
- Pyroptosis and necroptosis reporter cells
- Readily assessable HMGB1::Lucia reporter activity
- Guaranteed mycoplasma-free
Levels of IL-1β can be measured by Western blot, ELISA, or using InvivoGen’s HEK-Blue™ IL-1β cellular assay. Alternatively, secreted IL-18 can be detected using InvivoGen’s HEK-Blue™ IL-18 cellular assay.
Download our Practical guide on Inflammasomes
References:
1. Broz P. and Dixit V.M., 2016. Inflammasomes: mechanism of assembly, regulation and signalling. Nat Rev Immunol. 16:407-20.
2. Kovacs S.B. and Miao E.A., 2017. Gasdermins: effectors of pyroptosis. Trends Cell Biol. 27:673-84.
3. Grootjans S. et al., 2017. Initiation and execution mechanisms of necroptosis: an overview. Cell Death Differ. 24:1184-95.
Disclaimer: These cells are for internal research use only and are covered by a Limited Use License (See Terms and Conditions). Additional rights may be available.
SPECIFICATIONS
Specifications
Pyroptosis and necroptosis monitoring
Complete RPMI 1640 (see TDS)
Each lot is functionally tested and validated.
CONTENTS
Contents
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Product:THP1-HMGB1-Lucia™ Cells
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Cat code:thp-gb1lc
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Quantity:3-7 x 10^6 cells
- 1 ml of Zeocin® (100 mg/ml)
- 1 ml of Normocin® (50 mg/ml)
- 1 tube of QUANTI-Luc™ 4 Reagent (Lucia luciferase detection reagent)
Shipping & Storage
- Shipping method: Dry ice
- Liquid nitrogen
- Avoid repeated freeze-thaw cycles
Storage:
Caution:
Details
Pyroptosis is a consequence of inflammasome activation.
Typically, inflammasome activation is a two-step process. A first signal (‘priming’), provided by microbial molecules such as lipopolysaccharide (LPS), induces NF-κB-dependent expression of pro-IL1β. The second signal, provided by structurally unrelated microbial molecules (e.g. Nigericin toxin) or danger signals, triggers inflammasome multimerization. This leads to caspase-1 self-activation, proteolytic maturation of IL-1β and IL-18, and cleavage of Gasdermin D (GSDMD). Subsequent GSDMD pore formation at the cell membrane elicits a rapid cell death associated with the release of IL-1β, IL-18, and HMGB1 [1,2].
Necroptosis is mainly implicated in conditions of disease or infection and is triggered by the formation of a necrosome. The necrosome is induced upon Tumor Necrosis Factor (TNF) receptor or PRR activation in absence of caspase-8 and cIAP (cellular inhibitor of apoptosis) activity. The necrosome contains receptor-interacting serine/threonine-protein kinases RIPK1 and RIPK3, and the mixed lineage kinase-like MLKL. Phosphorylated MLKL relocalizes at the plasma membrane where it drives cell lysis and release of HMGB1, most probably through pore formation [3].
1. Broz P. and Dixit V.M., 2016. Inflammasomes: mechanism of assembly, regulation and signalling. Nat Rev Immunol. 16:407-20.
2. Kovacs S.B. and Miao E.A., 2017. Gasdermins: effectors of pyroptosis. Trends Cell Biol. 27:673-84.
3. Grootjans S. et al., 2017. Initiation and execution mechanisms of necroptosis: an overview. Cell Death Differ. 24:1184-95.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
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