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: Cell resuscitation steps and culture conditions
Cell resuscitation steps and culture conditions
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: What does the product contain?
What does the product contain?
Q8: Whether Puromycin needs to be continuously added during cultivation
Whether Puromycin needs to be continuously added during cultivation
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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