TransIT®-2020 Transfection Reagent

Figure 1. TransIT®-2020 Reagent Efficiently Transfects Human Induced Pluripotent Stem (iPS) Cells. The TransIT-2020 Transfection Reagent was used to transfect 0.5 x 10⁶ iPS cells with a ZsGreen expressing plasmid (Clontech). Transfections were performed in 6-well plates using 7.5 µl of TransIT-2020 Transfection Reagent to deliver 2.5 µg of DNA (3:1, reagent: DNA).  Cells were visualized 24 hours post-transfection and imaged at 4X objective with an Olympus IX71^® Inverted Microscope. Images are (A) phase contrast, and (B) green fluorescence. Cells were also assayed 24 hours post-transfection on an Accuri^® Cytometer. The histogram (C) shows untransfected cells (black line) compared to cells transfected with plasmid using TransIT-2020 (green line). See more information on stem cell applications.

Data courtesy of

Figure 2. TransIT®-2020 Reagent Exhibits Higher Expression and Lower Cellular Toxicity Compared to Other Transfection Reagents. Human umbilical vein endothelial cells (HUVEC) 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 (bar graph) and lactate dehydrogenase (LDH) levels (line graph) were measured at 24 hours post-transfection. LDH levels are reported as % cytotoxicity compared to cells alone and were measured using a commercially available colorimetric assay; all values at or below zero are represented as zero on graph. Error bars represent the standard deviation of triplicate wells.

Figure 3. Superior Gene Expression in a Broad Spectrum of Cell Types. The indicated cell types were transfected in 96-well plates with a luciferase expression plasmid (0.1ug/well) according to the manufacturer’s protocol. Reagent to DNA ratios were optimized for each cell type:  TransIT®-2020 (Mirus Bio, 2:1 or 3:1), FuGENE® HD (Roche, 3.5:1), Lipofectamine™ 2000 (Life Technologies, 1.5:1, 3:1 or 5:1).  Luciferase activity was measured 24 hours post-transfection. Values were normalized to TransIT-2020 and presented as a percentage of luciferase expression. FuGENE is a registered trademark of Fugent LLC. Lipofectamine is a trademark of Life Technologies Corporation.

Figure 4. TransIT®-2020 Reagent Effectively Transfects Drosophila S2 Cells. Cells were transfected with a plasmid construct expressing a secreted form of the Dscam extracellular domain fused to alkaline phosphatase (AP) in a 24-well plate. The ratio of transfection reagent to DNA and micrograms of DNA per well is noted beneath each bar. All products were used according to manufacturers’ protocol. All transfections were performed in serum-free media for four hours followed by complete media supplementation. An AP enzymatic assay was used to measure the AP levels 24 hours post-transfection.

Data courtesy of Woj Wojtowicz, University of California, Berkeley.