HEK-293T cells were obtained from American Type Culture Collection (ATCC) and were maintained in an incubator set at 37 °C and 5% CO2. They were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% inactivated fetal bovine serum (Gibco, Life Technologies, Thermo Fisher), 20 mM HEPES, 2 mM glutamine, 100 IU/ml penicillin, and 100 μg/ml streptomycin.
HEK-293 FreeStyle cells were obtained from ThermalFisher Scientific and were maintained in a New Brunswick S41i shaker (Eppendorf) set at 37 °C, 135 rpm, 8% CO2, and 100% humidity. They were cultured in SMM 293-TII media (Sinobiological), supplemented with 100 IU/ml penicillin and 100 μg/ml streptomycin.
BL21(DE3) and LOBSTR E. Coli strains were grown at 37 °C in a New Brunswick S44i shaker (Eppendorf) in LB media. To induce protein expression, they were transformed by the corresponding plasmids (see below), propagated at 37 °C, and induced with IPTG at 18 °C.
HEK-293 FreeStyle cells were obtained from ThermalFisher Scientific and were maintained in a New Brunswick S41i shaker (Eppendorf) set at 37 °C, 135 rpm, 8% CO2, and 100% humidity. They were cultured in SMM 293-TII media (Sinobiological), supplemented with 100 IU/ml penicillin and 100 μg/ml streptomycin.
BL21(DE3) and LOBSTR E. Coli strains were grown at 37 °C in a New Brunswick S44i shaker (Eppendorf) in LB media. To induce protein expression, they were transformed by the corresponding plasmids (see below), propagated at 37 °C, and induced with IPTG at 18 °C.