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Cat. No.: PIKN3-100 (for 100T)

Cat. No.: PIKN3-500 (for 500T)

Description

Pyroptosis Inducer Kit (NLRP3 Inflammasome Pathway) is a ready‑to‑use formulation combining lipopolysaccharide (LPS) and nigericin at an optimized ratio to robustly and reproducibly induce pyroptosis. By specifically activating the TLR4/NF‑κB signaling cascade and the NLRP3 inflammasome, the kit enables efficient, stable, and high‑fidelity induction of pyroptotic cell death. It is compatible with a wide range of monocyte and macrophage cell types and is suitable for mechanistic studies of pyroptosis, functional validation of related genes, and drug screening applications.

Pyroptosis is a form of programmed cell death mediated by the Gasdermin protein family and represents a key component of innate immune defense. It plays an essential role in combating infections and endogenous danger signals, while dysregulated pyroptosis is implicated in numerous diseases, including infectious disorders, autoinflammatory diseases, and cancer [1]. Cells undergoing pyroptosis typically exhibit pronounced swelling and membrane blebbing, followed by membrane rupture and release of intracellular contents, which in turn triggers a strong inflammatory response.

Gasdermin proteins contain a self‑inhibitory C‑terminal domain and an N‑terminal domain capable of forming membrane pores. During pyroptosis, Gasdermins are specifically cleaved, releasing the N‑terminal fragment that oligomerizes on the plasma membrane to form pores, disrupt osmotic balance, and cause membrane rupture. This process promotes the release of damage‑associated molecular patterns (DAMPs), including pro‑inflammatory cytokines such as IL‑1β and IL‑18, as well as LDH, ATP, and HMGB1.

Among the Gasdermins, Gasdermin D–mediated pyroptosis is the most extensively studied. Its activation occurs through two major pathways:

• Classical pathway: Triggered by pathogen‑associated molecular patterns (PAMPs) or DAMPs, the NLRP3 inflammasome recruits the adaptor ASC to activate Caspase‑1. Activated Caspase‑1 cleaves Gasdermin D to release its N‑terminal pore‑forming domain and simultaneously processes pro‑IL‑1β and pro‑IL‑18 for secretion, ultimately leading to membrane rupture and a potent inflammatory response.

• Non‑classical pathway: Human Caspase‑4/5 or murine Caspase‑11 directly recognize cytosolic LPS, cleaving Gasdermin D to induce osmotic imbalance and pyroptosis [2, 3]. The non‑classical pathway can also indirectly activate the NLRP3 inflammasome, amplifying downstream inflammatory signaling.

Cell death is fundamental to multicellular organism development, tissue homeostasis, immunity, and host defense. Regulated forms of cell death are collectively referred to as programmed cell death, whereas unregulated cell death is termed necrosis. Programmed cell death encompasses apoptosis, necroptosis, pyroptosis, ferroptosis, cuproptosis, and others.

LPS is a lipid–polysaccharide complex and a major structural component of the outer membrane of most Gram‑negative bacteria, exhibiting strong immunogenicity. LPS primarily binds to LPS‑binding protein (LBP) and subsequently forms a complex with Toll‑like receptor 4 (TLR4), initiating downstream signaling through the MyD88 pathway to induce inflammatory cytokine production. By activating the NF‑κB signaling cascade, LPS upregulates transcription and expression of pro‑inflammatory mediators and plays a critical role in priming cells for pyroptosis induction.

Nigericin is an antibiotic and a potent NLRP3 activator that triggers assembly of the NLRP3 inflammasome, leading to downstream Caspase‑1 activation and pyroptotic cell death. In addition, nigericin facilitates transmembrane K⁺/H⁺ exchange, disrupts Golgi apparatus function, and inhibits the growth of Gram‑positive bacteria.

This kit provides high efficiency, stability, and strong specificity. The optimized formulation enables robust induction of classical NLRP3 inflammasome–mediated pyroptosis with excellent reproducibility and minimal off‑target effects.

The kit is compatible with a broad range of monocyte and macrophage cell types. It effectively induces NLRP3 inflammasome–dependent pyroptosis in murine cells such as BMDMs, iBMDMs, Raw264.7‑ASC, and J774A.1, as well as human cells including differentiated THP‑1, U937, and hMDMs.

Storage Conditions

• –80°C: Stable for 1 year.
• –20°C: Stable for at least 3 months.
• 4°C: Stable for at least 3 days.

Precautions

• To ensure optimal performance, it is recommended to aliquot the reagent after the first thaw to avoid repeated freeze–thaw cycles.
• This product is intended for monocytes, macrophages, and other cell types capable of undergoing NLRP3 inflammasome–mediated pyroptosis.
• Because pyroptosis induction is highly cell‑type‑specific and the underlying mechanisms vary across cell models, this product cannot guarantee successful induction of pyroptosis in every cell type.
• This product is harmful to humans. Handle with care and use appropriate protective measures to avoid direct contact or inhalation.
• This product is for research use only by trained professionals. It is not intended for clinical diagnosis or therapy, not for use in food or pharmaceuticals, and must not be stored in residential settings.
• For your safety, wear a lab coat and disposable gloves during operation.