For transfection, HEK293T cells were cultured to the concentration of 30%. Transfection was performed in 500 μl opti-DMEM with 20 μl siRNA or control with addition of 30 μl X-tremeGene (Roche, Mannheim, Germany) according to the manufacturer’s instruction. 2 μg constructions for ectopic expression of CUL4B, IRGM1 and HA-Ubiquitin were transfected by PEI transfection reagent (Merck) as indicated respectively. After 48 h incubation, MG132 was added at a final concentration of 10 μM. The cells were then harvested after 3 h incubation.
Plent ef1a fh cmv gfp p2a puro
PLent-EF1a-FH-CMV-GFP-P2A-Puro is a lentiviral vector that contains the elongation factor 1-alpha (EF1a) promoter, a FLAG-HA (FH) tag, the cytomegalovirus (CMV) promoter, green fluorescent protein (GFP), a P2A self-cleaving peptide, and a puromycin resistance gene (Puro). This vector can be used for the expression and monitoring of proteins of interest in transduced cells.
Lab products found in correlation
3 protocols using plent ef1a fh cmv gfp p2a puro
Lentiviral Expression of IRGM1 and CUL4B
For transfection, HEK293T cells were cultured to the concentration of 30%. Transfection was performed in 500 μl opti-DMEM with 20 μl siRNA or control with addition of 30 μl X-tremeGene (Roche, Mannheim, Germany) according to the manufacturer’s instruction. 2 μg constructions for ectopic expression of CUL4B, IRGM1 and HA-Ubiquitin were transfected by PEI transfection reagent (Merck) as indicated respectively. After 48 h incubation, MG132 was added at a final concentration of 10 μM. The cells were then harvested after 3 h incubation.
Lentiviral Vector Production of BPV-13 E5
Lentiviral Vector Production Protocol
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