HEK-293FT (Thermo Fisher Scientific) cells were cultured in DMEM (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS), 2 mM Glutamine, 100 units/ml Penicillin, 100 μg/ml streptomycin sulfate, and 1 mM sodium pyruvate (Thermo Fisher Scientific). The HEK-293FT cells were plated at a cell dose of 1 × 107 on a 10-cm tissue culture dish and incubated at 37°C 5% CO2 incubator until the cells reach 80% confluence. The HEK-293FT cells were cotransfected either lentiviral ORFs, the constructs shown in Figure 1a , along with Lenti-vpak Packaging Kit (OriGene) using the transfection reagent MegaTran 1.0 (OriGene). After 48–72 h transfection, the virus-containing medium was collected, spun down, filtered (0.45 μm), and used for targeting into NCI-H82 by infection. The virus-infected stable clones were obtained after at least 2–3 weeks of selection in 10% FBS/RPMI-1640 with 0.5 μg/ml of Puromycin (Sigma-Aldrich). Protein expression in stable clones was confirmed by Western blot analysis.
Hek 293ft
Manufactured by OriGene
HEK-293FT is a widely used human embryonic kidney cell line. It is a highly transfectable variant of the HEK-293 cell line, suitable for the production of lentiviral and retroviral vectors.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (3 mentions)
Lenti vpak packaging kit, by OriGene (2 mentions)
Puromycin, by Merck Group (2 mentions)
Megatran 1, by OriGene (2 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (2 mentions)
Streptomycin sulfate, by Thermo Fisher Scientific (2 mentions)
Hek293ft, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
2 protocols using hek 293ft
1
Lentiviral Transduction of HEK-293FT Cells
2
Lentiviral Transduction of Lymphoid Cells
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HEK293FT (Life Technologies) cells were cultured in Dulbecco's modified Eagle's medium (Thermo Fisher Scientific) supplemented with 10% FBS, 2 mM glutamine, 100 units/ml penicillin, 100 µg/ml streptomycin sulfate and 1 mM sodium pyruvate (Life Technologies). Ten million cells were plated at cell on a 10 cm tissue culture dish and incubated at 37 °C 5% CO2 incubator until the cells reach 80% confluence. The HEK293FT cells were co-transfected with either lentiviral open reading frames or shRNAs along with Lenti-vpak Packaging Kit (OriGene) using the transfection reagent MegaTran 1.0 (OriGene). After 48–72 h of transfection, the virus-containing medium was collected, spun down, filtered (0.45 µm) and used for targeting into CCRF-CEM or MOLT-4 cells by infection. The virus-infected stable clones were obtained after at least 2–3 weeks of selection in 10% FBS/RPMI-1640 with 0.5 µg/ml of puromycin (Sigma-Aldrich).
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