Contents and Storage

? 3-7 x 106 A549-ASC cells in a cryovial or shipping flask IMPORTANT: If cells provided in a cryovial are not frozen upon arrival,  contact InvivoGen immediately.

? 1 ml of Blasticidin(10 mg/ml). Store at 4°C or at -20°C.*

? 1 ml of Normocin™ (50 mg/ml), a formulation of three antibiotics active against mycoplasmas, bacteria and fungi. Store at -20°C.*

*The expiry date is specified on the product label. Handling Frozen Cells Upon Arrival Cells must be thawed immediately upon receipt and grown according  to handling procedures (as described on the next page) to ensure the  best cell viability and proper assay performance. Note: Avoid freezing cells upon receipt as it may result in irreversible  damage to the cell line.

Disclaimer: We cannot guarantee cell viability if the cells are not thawed  immediately upon receipt and grown according to handling procedures. IMPORTANT: For cells that arrive in a shipping flask please refer to the  enclosed ‘cell recovery procedure’.

Cell Line Stability

Cells will undergo genotypic changes resulting in reduced  responsiveness over time in normal cell culture conditions. Genetic  instability is a biological phenomenon that occurs in all stably  genetically engineered cells. Therefore, it is critical to prepare an  adequate number of frozen stocks at early passages. To ensure  maximum efficiency, do not passage A549-ASC cells more than 20  times.

Quality Control

? The induction of ASC::GFP expression has been verified by Western  blot, fluorescence microscopy and flow cytometry.

? The stability of this cell line for 20 passages following thawing has been verified.

? A549-ASC cells are guaranteed mycoplasma-free.

Applications:

• Screening of NLRP1 inflammasome activating stimuli

• Real-time monitoring of ASC-dependent inflammasome assembly

• Rapid and visual screening of drugs targeting the NLRP1 inflammasome by fluorescence microscopy

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