Lipofectamine® 2000 Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Do not add antibiotics to the Transfection Medium

Protocol tips
Do not add antibiotics to the Transfection Medium

Publication protocol

Transient transfection and chloramphenicol acetyl transferase (CAT) assay
In general, 5 × 106 HSCs were transiently transfected before confluence by the LipofectAMINE technique (Invitrogen, Carlsbad, CA) (17). Cell lysates were prepared, and CAT activity was determined as described elsewhere (19). Luciferase activity was determined using the enhanced luciferase assay kit according to the manufacturer's protocol (Analytical Luminiscence, San Diego, CA). Cell lysates were prepared in 125 μl of cell lysis buffer. Luciferase activity was determined using 50 μl aliquots, and protein concentrations determined with 5-μg aliquots using the Bradford protein assay (Bio-Rad Laboratories, Hercules, CA) (20). All reporter genes were normalized for transfection efficiency by cotransfecting a constant amount of the pRSVβ-gal reporter gene and determining β-galactosidase activity (21).



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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Lipofectamine® 2000 Transfection Reagent below.

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