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Contents

• 3-7 x 106 A549-Dual™ hACE2-TMPRSS2 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 Hygromycin B Gold (100 mg/ml). Store at 4°C or at -20 °C.* • 1 ml of Puromycin (10 mg/ml). Store at 4°C or at -20 °C.*

• 1 ml of Zeocin® (100 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.

• 1 tube of QUANTI-Luc™ 4 Reagent, a lucia luciferase detection reagent (sufficient to prepare 25 ml). Store at -20°C. Avoid repeated freeze-thaw cycles. Note: This product is photosensitive and should be protected from light.

• 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent). Store QB reagent and QB buffer at -20°C. QUANTI-Blue™ Solution is stable for 2 weeks at 4°C and for 2 months at -20°C. Note: Data sheets for all components are available on our website. 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 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 transfected cells. Therefore, it is critical to prepare an adequate number of frozen stocks at early passages. A549-Dual™ hACE2-TMPRSS2 cells should not be passaged more than 20 times to remain fully efficient.

Quality Control

• The overexpression of the human ACE2 (hACE2) gene has been verified by RT-qPCR, FACS staining, and functional assays.

• The overexpression of the human TMPRSS2 (hTMPRSS2) gene has been verified by RT-qPCR and functional assays.

• Lucia luciferase and SEAP reporter activities have been validated using functional assays.

• The stability for 20 passages following thawing has been verified.

• These cells are guaranteed mycoplasma-free.