TransIT®-LT1 Transfection Reagent

A broad spectrum, low toxicity, DNA transfection reagent

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  • Broad Spectrum DNA Delivery – Utilize one transfection reagent and protocol for a variety of cells
  • Low Cellular Toxicity – Maintain cell density and reduce experimental biases  Learn more >>
  • High Efficiency Delivery – Achieve expression in a large population of cells for experimental success
  • Deliver Single or Multiple Plasmids – Suitable for applications such as virus production

NEW PROTOCOL! Reverse Transfection of Human Induced Pluripotent Stem (iPS) Cells with TransIT®-LT1 Transfection Reagent

Figures and Data

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TransIT-LT1 Reagent Efficiently Transfects Human Induced Pluripotent (iPS) Cells

Exceptional Transfection Efficiency in Human Induced Pluripotent Stem Cells (iPSCs) via Reverse Transfection with TransIT®-LT1. The TransIT®-LT1 Transfection Reagent was used to reverse transfect 1.3 x 106 iPS cells with a ZsGreen expressing plasmid (Clontech). Reverse transfections were performed in 6-well plates using 12 µl of TransIT®-LT1 Transfection Reagent to deliver 4 µg of DNA (3:1, reagent: DNA). Cells were visualized 48 hours post-transfection and imaged under a 10X objective with an Olympus IX71® Inverted Microscope. Images are (A) phase contrast and (B) green fluorescence. Cells were assayed 48 hours post-transfection on an Accuri® Cytometer. The histogram (C) shows untransfected cells (black line) compared to cells transfected with plasmid using TransIT®-LT1 (green line). See more information on stem cell applications.

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High Efficiency Transfection of iCell Cardiomyocytes Using TransIT-LT1

High Efficiency Transfection of iCell® Cardiomyocytes Using TransIT®-LT1 Transfection Reagent. iCell® Cardiomyocytes were plated at 20,000 cells/well in a 96 well tissue culture plate coated with 0.1% gelatin. After allowing the cells to recover from thaw, cells were transfected with 100 ng/well of pMAXGFP (Lonza) using TransIT®-LT1 Transfection Reagent with a 2:1 reagent-to-DNA ratio according to the manufacturer’s instructions. Fluorescent images were taken 3 days post transfection using a Olympus IX71® inverted microscope. See more information on stem cell applications.

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Higher Expression and Lower Toxicity with TransIT-LT1

The TransIT®-LT1 Reagent Exhibits Higher Expression and Lower Cellular Toxicity Compared to Other Transfection Reagents. HepG2 cells were transfected with a luciferase expression plasmid using the designated reagents at the manufacturer’s recommended reagent-to-DNA ratio 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. Experiments were performed as per industry accepted testing protocols. FuGENE® is a registered trademark of Fugent LLC. Lipofectamine® is a trademark of Life Technologies Corporation.

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Comparable Luciferase Expression with TransIT-LT1 and FuGENE 6 in Multiple Cell Types

Comparable Luciferase Expression with the TransIT®-LT1 Reagent and FuGENE® 6 in Multiple Cell Types. The indicated cell lines were transfected in duplicate with 1 µg of a luciferase expression vector per well of a 12-well plate using either 3 µl of the TransIT®-LT1 or FuGENE® 6 Reagents according to industry accepted testing protocols. Cells were harvested 24 hours post-transfection and assayed for luciferase activity. FuGENE® is a registered trademark of Fugent LLC.

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Specifications

Storage Conditions:
All Configurations: Store at 4°C

Product Guarantee:
All Configurations: 1 year

Usage Statement:
All Configurations: For Research Use Only.

Animal Origin Statement:
All Configurations: This product contains raw materials that are derived from animals.

Technical Product Literature


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Testimonials

We routinely use Mirus TransIT®-LT1 Transfection Reagent for the delivery of plasmid DNA to carry out immunoprecipitation experiments. Our lab recently published using TransIT®-LT1 for this application to reveal a crucial regulator (MCUR1) for calcium uptake in the mitochondria to regulate cellular metabolism. (Mallilankaraman, K et al. Nature Cell Biology. December 2012)
Dr. Karthik Mallilankaraman
Madesh Lab, Temple University
Our lab is interested in identifying barriers to influenza virus cross-species transmission and how those control the emergence of new pandemic viruses... we were able to use TransIT®-LT1 in our studies to further implicate the viral polymerase complex as a major regulator of viral tropism and identify new adaptive strategies used by influenza as it moves from birds to people. Details of these efforts can be found in the Journal of Virology. (Mehle, A et al. Journal of Virology. Feb 2012)
Dr. Andrew Mehle
University of Wisconsin - Madison
Our lab regularly transfects head and neck cancer squamous carcinoma cell lines as well as pre-cancerous oral lesion cell lines and primary cells with Mirus Bio products. We recently published a paper using Mirus Bio TransIT®-LT1 in which we concluded tobacco carcinogens up-regulate AP-1 activity and AP-1 dependent IL-8 and VEGF gene expression in head and neck cancer. (Swenson, W et al. Molecular Carcinogenesis. April 2011)
Beverly Wuertz
Ondrey Lab, University of Minnesota
I recently tested TransIT®-2020 and TransIT®-LT1, and both reagents worked well in terms of their efficiency at transfecting human-derived iPS cells with CRISPR constructs and a fluorescent protein reporter. Through visual inspection, transfection efficiencies with TransIT®-2020 and TransIT®-LT1 were clearly higher than with Lipofectamine® 3000.
Fedir Kiskin
University of Cambridge
Our lab tested TransIT®-LT1 from Mirus for virus production in 293T cells. At a 3:1 ratio using up to 5 µg of lentivirus, packaging and envelope plasmid, TransIT®-LT1 was far superior to Lipofectamine® in terms of viral titer downstream. So much so that our lab purchased the 0.4 ml product after using the trial.
Andrew Paluch
University of Cincinnati