TransIT®-BrCa Transfection Reagent

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.

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.

TransIT®-BrCa Transfection Reagent Yields High Efficiency Plasmid DNA Transfection. 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 24-well plates using 1-1.5 µl of TransIT®-BrCa Transfection Reagent to deliver 0.5 µg of DNA (2:1 and 3:1, reagent:DNA ratio). Triplicate wells were assayed 48 hours post-transfection on a Guava® HT easyCyte™ flow cytometer.

Activation of Estrogen Receptor Pathway is Detected in MCF-7 Cells Transfected with TransIT®-BrCa Transfection Reagent. MCF-7 (estrogen receptor positive) and MDA-MB-231 (estrogen receptor negative) breast cancer cells were maintained in complete media containing charcoal stripped FBS for 3 days prior to transfection. TransIT®-BrCa Transfection Reagent was used to deliver an ERE-luciferase reporter plasmid or negative control (SA Biosciences) at a 2:1 reagent-to-DNA ratio. Transfections were performed in 96-well plates using 0.1 μg of plasmid DNA per well. Twenty four hours post-transfection, cells were treated with varying levels of 17-estradiol (E2) for 6 hours. Firefly and Renilla luciferase expression was measured at 30 hours post-transfection using a standard assay. Promoter activity is represented as the ratio of firefly to luciferase relative light units (RLU) using the Renilla luciferase for normalization. Error bars represent the standard deviation of triplicate wells.