( sity ofa reporter gene and cellular ...)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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