To establish stable doxycycline-inducible cell lines expressing either FLAG-tagged tail-anchored proteins or FLAG-tagged ATP13A1, wildtype or ATP13A1 knockout Flp-In T-REx HeLa or 293 cells were co-transfected with a 1:1 ratio of pOG44 and pcDNA5/FRT/TO vector containing the desired insert using Lipofectamine 2000/Lipofectamine 3000 or TransIT293 (Mirus Bio), respectively. After 24 hr, cells were selected with 10 μg/mL Blasticidin and 150-300 μg/mL Hygromycin for at least two weeks, and expression was validated by induction with 10 ng/mL doxycycline for 24-48 hr and immunoblotting.
Transit 293
TransIT-293 is a transfection reagent designed for the efficient delivery of nucleic acids, such as plasmid DNA, into 293 and 293T cell lines. It is a proprietary, non-liposomal formulation that facilitates the uptake of DNA into cells, enabling effective gene expression.
Lab products found in correlation
155 protocols using transit 293
Generation and Validation of Inducible Cell Lines
Transient and Stable Transfection of HEK293 Cells
Recombinant Antibody Production in 293T Cells
Lentiviral Transduction and RNAi Knockdown
Overexpression and Detection of scAZ1-avitag
Overexpression and Detection of scAZ1-avitag
Transient AAV Vector Expression in 293T Cells
Example 8
Each individual AAV expression vectors were transfected into 293T cells with TransIT-293 (Mirus, Inc.) to test their function before being packaged into AAV viral vectors. 293T cells were transfected with the same amount of plasmid and harvested at the same time points. SaCas9 protein expression was assessed by western blotting with primary antibody probing for the triple Flag tag at the C-terminus of saCas9, while loading control was demonstrated by αTubulin expression. Deletion events at IVS26 mutation could be determined by PCR amplification followed by Sanger sequencing or ddPCR. The results are shown in
Reverse Genetics for Influenza A Virus
Production and Characterization of HIV Variants
Dual-Luciferase Assay for Transfection Efficiency
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