Study activation of ASC-dependent inflammasomes in real time.
Canonical inflammasomes  require the adaptor protein ASC (apoptosis-associated speck-like protein with a CARD; also known as PYCARD) for the activation of caspase-1. After inflammasome activation, ASC assembles into a large protein complex, which is termed "speck". In most cells only one speck forms upon inflammasome activation [2, 3].
THP1 ASC-GFP cells stably express an ASC::GFP fusion protein that enables the visual monitoring of ASC specks. These cells are derived from THP-1 human monocytic cells, which are the most commonly used model cell line for the study of inflammasome activation. The expression of ASC::GFP is driven by an NF-kB-inducible promoter. The assembly of the ASC-dependent inflammasome requires a priming signal from a pathogen-associated molecular pattern (PAMP), such as lipopolysaccharide (LPS) that induces NF-kB and the subsequent production of ASC::GFP. The second signal provided by an inflammasome inducer such as nigericin promotes the formation of ASC::GFP specks. These fluorescent specks can be visualized by using time-lapse confocal or high-resolution fluorescence microscopy. The number and localization of ASC::GFP positive cells can be determined and provide information on inflammasome formation.
Antibiotic resistance: Zeocin
Growth medium: RPMI 1640 (2 mM L-glutamine, 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES and 1.0 mM sodium pyruvate) with 10% heat-inactivated fetal bovine serum, 100 µg/ml Normocin™, Pen-Strep (50 U/ml-50 μg/ml)
Quality control: The functionality of THP1 ASC-GFP cells has been tested using inflammasome inducers, such as the microbial toxin nigericin and transfected poly(dA:dT). The stability of this cell line for 20 passages following thawing has been verified. THP1 ASC-GFP cells are guaranteed mycoplasma-free.
Shipped on dry ice.
This product is covered by a Limited Use License (See Terms and Conditions).
• 1 vial of THP1 ASC-GFP cells (3-7 x 106 cells) in Freezing Medium.
• 1 ml Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria and fungi.
• 100 μl Zeocin (100 mg/ml).
- Broz P. & Dixit V., 2016. Inflammasomes: mechanism of assembly, regulation and signalling. Nat Rev Immunol. 16:407-20.
- Dick M. et al., 2016. ASC filament formation serves as a signal amplification mechanism for inflammasomes. Nat Commun. 7:11929.
- Stutz A. et al., 2013. ASC speck formation as a readout for inflammasome activation. Methods Mol Biol. 1040:91-101.