TransIT^® Cell Line Specific Transfection Reagents

Fig. 1 | TransIT^®-293 Reagent Achieves High Transfection Efficiency. HEK 293 cells transfected with pEGFP using TransIT^®-293 Reagent in complete media for 24 hours.

Fig. 2 | TransIT^®-BrCa Transfection Reagent Exhibits Higher Luciferase Expression in Breast Cancer Cells Compared to Other Transfection Reagents. Breast Cancer cell lines, MCF-7, MDA-MB-231, MDA-MB-453, MDA-MB-468 and T47D, were transfected with a luciferase expression plasmid using the designated reagents at the reagent-to-DNA ratios indicated beneath each bar. Transfections were performed in 96-well plates using 0.1 µg of plasmid DNA per well. Luciferase expression was measured at 24 hours post-transfection using a standard assay. Error bars represent the standard deviation of triplicate wells.

Fig. 3 | High Efficiency Delivery of Plasmid DNA in Breast Cancer Cell Types. TransIT^®-BrCa Transfection Reagent was used to transfect plasmid DNA encoding GFP into MCF-7, MDA-MB-231, MDA-MB-453, MDA-MB-468 and T47D breast cancer cell lines. Transfections were performed in 35 mm MatTek dishes using 4 µl of TransIT^®-BrCa Transfection Reagent to deliver 2 µg of DNA (2:1 reagent:DNA ratio). Images (32X) were captured at 48 hours post-transfection using a Zeiss Axiovert S100 inverted fluorescence microscope.

Fig. 4 | TransIT^®-Keratinocyte Reagent Achieves High Transfection Efficiency. Near-Diploid Immortalized Keratinocytes transfected with a Beta-galactosidase reporter vector using TransIT^®-Keratinocyte Reagent in complete media for 48 hours. Reporter protein was detected using Mirus Bio’s Beta-Galactosidase Staining Kit.

Fig. 5 | TransIT^®-Insect Outperforms Competitor Transfection Reagents. Insect cell lines (A) Sf9, (B) High Five™, and (C) Drosophila S2 cells were transfected in 96-well plates with 0.1 µg of a luciferase expression plasmid driven by an hr5 enhancer/IE1 promoter using the designated reagent at the indicated reagent-to-DNA ratios (µl:µg). Luciferase expression was measured at 48 hours post-transfection. Sf9 and High Five™ cells were cultured and transfected in serum-free media formulations; S2 cells were in serum containing medium. Error bars represent the standard error of the mean for triplicate wells.

Fig. 6 | Superior Recombinant Protein Expression in High Five™ Cells Using TransIT^®-Insect. High Five™ cells were transfected in 6-well plates with 2.5 µg of a GFP expression plasmid driven by an hr5 enhancer/IE1 promoter using the designated reagent at the indicated reagent-to-DNA ratios (µl:µg). Total soluble cell lysates were prepared from cells 72 hours post-transfection. Lysates from 100 µl culture were analyzed by SDS-PAGE and Coomassie blue staining; cells alone (untransfected) is shown as control. Expressed GFP containing 6X His, S, and HSV tags (~38 kDa) was clearly detected in the lysate from the cells that were transfected (*) with the highest level of expression observed at TransIT^®-Insect:DNA ratio of 2:1.

Fig. 7 | TransIT^®-Insect Yields Increased Protein Expression Over Time. Insect cell lines (A) Sf9, (B) High Five™, and (C) Drosophila S2 were transfected in a 96-well plate with 0.1 µg of a luciferase expression plasmid driven by an hr5 enhancer/IE1 promoter using the TransIT^®-Insect Transfection Reagent at a reagent-to-DNA ratio of 2:1 (µl:µg). Luciferase expression was measured at three time points, 24, 48 and 72 hours post-transfection. Sf9 and High Five™ cells were cultured and transfected in serum-free media formulations; S2 cells were in serum containing medium. Error bars represent the standard error of the mean for triplicate wells.