Q1: What does the product include?
The two cells we provide are: (1) Negative cells = blank vector control +puro (that is, cells infected with empty virus, which gives the cells the ability to grow in a puro environment) (2) Positive cells = 293 cells transferred to ACE2 +puro
Q2: Does the culture medium need to add antibiotics?
Double antibiotics can be omitted. However, if the culture environment is rather complex, it is recommended to add double antibiotics to avoid contamination. The function of puromycin: Screen 293T cells with overexpression of ACE2 and inhibit the growth of negative cells without overexpression of ACE2. High-concentration drug maintenance (purinomycin) has an impact on cell growth
Q3: Shipped as cryovials or T25 flasks?
Depending on the customer's requirements, if they want to freeze the storage, we will ship the cryotubes. If they want to revive, we will ship T25 bottles. By default, cryotubes will be shipped
Q4: Is Puromycin needed throughout the culture?
Subculture 1-2 times and then add the drug for screening. There is no need to maintain it all the time. High-concentration drug maintenance also has an impact on cell growth
Q5: 细胞复苏步骤及培养条件
复苏：37℃水浴；培养条件：10%FBS+DMEM；Puromycin浓度：5μg/ml
Q6: Has it been validated by flow cytometry?
No, we also have a monoclonal ACE2 stable cell line, which was prepared by the limited dilution method and finally verified by viral infection. Some customers have constructed 293t cells of ACE2-mcherry. The flow cytometry they took back was fine, but we don't have experimental data.
Q7: 产品包含什么
阴性细胞=空白载体对照+puro；阳性细胞=转入ACE2的293细胞+puro
Q8: 培养是否需要一直加Puromycin
传代1-2次加药筛一下就可以，不需要一直维持
Q9: Cell thawing steps and culture conditions
Resuscitation: 37℃ water bath; Cultivation conditions: 10%FBS+DMEM; Puromycin concentration: 5μg/ml
Q10: Has it been validated by flow cytometry?
There was no flow cytometry verification. The verification method was virus infection verification
Q11: Can experimental data for ACE2 stable cell lines be provided?
There are no experimental data on ACE2 stable cell lines
Q12: Can 293T cells be used for virus amplification?
Sure, 293T cells can be used for virus amplification, but vectors carrying genes related to virus amplification, such as pCD513B1, are required.
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