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| Product Name | HEK293T-ACE2 Overexpression Cell Line |
| Catalog No. | RESK-0606 |
| Description | ACE2 (angiotensin-converting enzyme 2) is a type I integral membrane protein widely distributed in tissues such as lung, heart and kidney, where it regulates blood pressure and fluid balance. It also serves as a functional receptor for coronaviruses, mediating viral entry and cell fusion. ACE2 consists of an N-terminal peptidase domain (PD) and a C-terminal collectrin-like domain (CLD); the extracellular PD region binds the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. With the aid of specific proteases, the spike protein can induce shedding of the ACE2 extracellular region, while the intracellular domain of ACE2 assists viral entry and infection. This cell line is a stable line generated by infecting 293T cells with an ACE2-overexpressing lentivirus followed by selection; it overexpresses human ACE2. |
| Intended Use | Cell model for coronavirus entry studies, including SARS-CoV-2 spike protein pseudovirus infection assays, neutralizing antibody research and antiviral screening. |
| Principle / Technology | Stable overexpression of human ACE2 on the surface of HEK293T cells, established by lentiviral transduction and antibiotic (puromycin) selection, rendering the cells highly permissive to spike-protein-mediated pseudovirus entry. |
| Detection Method | Trypan blue staining for viable cell counting; pseudovirus infection can be read out with a suitable reporter gene assay. |
| Sample Type | Cryopreserved human HEK293T-derived stable cell line, 5 x 10^6 cells/mL, 1 mL per vial. |
| Performance Range / Specifications | Cryopreserved at 5 x 10^6 cells/mL in 1 mL per vial. Typical adherent cell densities: 35 mm dish 3.2 x 10^5 (confluent 1.3 x 10^6); 60 mm dish 7.8 x 10^5 (confluent 3.1 x 10^6); 100 mm dish 2.3 x 10^6 (confluent 9.0 x 10^6); T-25 flask 6.9 x 10^5 (confluent 2.6 x 10^6); T-75 flask 2.1 x 10^6 (confluent 8.4 x 10^6). |
| Specificity | Overexpresses human ACE2; derived from human HEK293T cells. |
| Reaction Conditions / Protocol | Cell recovery: thaw the vial rapidly in a 37 °C water bath; transfer the cell suspension into 9 mL pre-warmed complete medium, centrifuge at 800 rpm for 5 min, discard the supernatant and resuspend in 15 mL medium (viable cell density should be 2 x 10^5 to 3 x 10^5 cells/mL). Seed into one 10 cm dish (10 mL) and one 6 cm dish (5 mL) at 20-30% density and incubate at 37 °C for 24 h; the first passage can usually be performed about 48 h after recovery. Passaging: use 0.25% Trypsin-EDTA for 30-60 s at 37 °C; stop digestion with about 2 mL complete medium, centrifuge at 800 rpm for 5 min at room temperature and reseed. Recommended split ratio 1:4 to 1:5 every other day; passage when confluence exceeds 60% and do not allow confluence to exceed 80%. Cryopreservation: collect cells by centrifugation at 1000 rpm for 3 min, resuspend at 2 x final density in pre-cooled FBS, mix with an equal volume of 2 x freezing medium (80% FBS + 20% DMSO), aliquot 1 mL per cryovial at 5 x 10^6 cells/mL, place in a controlled-rate freezing container at -80 °C for at least 1 day, then transfer to liquid nitrogen as soon as possible. |
| Components / Formulation | Recommended media (not included): Recovery medium: DMEM + 10% FBS + 1% penicillin-streptomycin Growth medium: DMEM + 10% FBS + 1% penicillin-streptomycin + 0.75 μg/mL puromycin Freezing medium: 90% FBS + 10% DMSO |
| Storage Conditions | Store in liquid nitrogen if the cells are not recovered immediately upon receipt; confirm that the product is still frozen on arrival. |
| Package Specifications | 1 mL (5 x 10^6 cells/mL) |
| Product Form | Cryopreserved cells |
| Quality Control | Stable cell line generated by lentiviral transduction of 293T cells followed by selection. Quantitative ACE2 protein content and passage-stability testing have not been performed. |
| Concentration | 5 x 10^6 cells/mL |
| Source / Origin | Human HEK293T cells stably overexpressing human ACE2 |
| Shipping Conditions | Shipped on dry ice. |
| Expiration Date / Stability | Maintain in liquid nitrogen for long-term storage; avoid repeated freeze-thaw cycles. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Suitable for pseudovirus infection assays and reporter gene readouts (e.g., luciferase-based pseudovirus systems). |
| Application Notes / Precautions | Upon receipt, immediately confirm that the product is still in a frozen state; if not recovering immediately, transfer to liquid nitrogen at once. Wipe the outside of the cryovial with 70% ethanol before opening and handle in a biosafety cabinet or clean bench. Passage cells when confluence is greater than 60% and never allow confluence to exceed 80%. If cells are not fully attached 24 h after recovery, continue incubation up to 48 h. Avoid contamination with bacteria, fungi or mycoplasma during culture. |
For research use only, not for clinical use.
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