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Versatile Transfection Reagent Offers Low Toxicity and Consistent,,,Performance

sity of a reporter gene and cellular protein concentration across a matrix of conditions. Using this approach, we performed several comparison studies to assess transfection efficiency and cellular toxicity, then determined that LipoTAXI transfection reagent consistently outperformed other lipid formulations.3,5

Optimizing Transfection Conditions

To perform convenient optimization studies, researchers can use the control plasmid included with LipoTAXI transfection reagent. This plasmid encodes the b-galactosidase enzyme regulated by the cytomegalovirus (CMV) promoter. We used this plasmid DNA (0.5 to 1.5 g) and LipoTAXI reagent (5 to 12.5 l), complexed together in serum-free media, to transfect 1 x 105 prostate epithelial cells (PrEC) from Clonetics Corporation (Figure 1A). Transfection conditions, preparation of cell lysates, and measurement of b-galactosidase activity were performed as previously described.3 In these experiments, increased b-galactosidase activity resulted from using higher DNA concentrations. However, as the concentration of DNA increased, cellular protein levels decreased (Figure 1B), indicating cytotoxicity. Cell death, associated with cytotoxicity, excludes a subpopulation of cells from transfection studies and should be avoided for results to be considered physiologically relevant. High protein content, which is associated with vigorous cell growth, is preferred.

Table 1
Optimized Concentrations of DNA and LipoTAXi'"/>

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