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TransIT-TKO® Transfection Reagent

A high efficiency, low toxicity, siRNA and plasmid DNA transfection reagent for mammalian cells

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  • Broad Spectrum siRNA and Plasmid DNA Delivery – Utilize one transfection reagent and protocol for a wide variety of cells
  • Low Cellular Toxicity – Maintain cell density and reduce experimental biases
  • Reproducible Results – Obtain consistent, targeted gene knockdowns from day to day
  • High Knockdown Efficiency – Achieve optimal gene silencing in a large percentage of cells to ensure experimental success

"We have tried other transfection reagents, but only the TransIT-TKO® reagent gives us a 100% transfection rate and gene knockdown without toxicity in these cells (RAW 264.7)." - Nature Protocols 1: 508 - 517 (2006)

Additional Information

NEW! Stem cell applications
NEW! Optimized transfection protocol for iCell® Cardiomyocytes (Cellular Dynamics International) using TransIT-TKO® Transfection Reagent: PROTOCOL | iCell® Page
Cell Types successfully transfected at Mirus using TransIT-TKO® Transfection Reagent

Figures and Data

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High Efficiency Endogenous Knockdown in iCell Cardiomyocytes

High Efficiency Endogenous Knockdown in iCell® Cardiomyocytes. The TransIT-TKO® Transfection Reagent was used to transfect iCell® Cardiomyocytes (Cellular Dynamics International) plated at a density of 136,500 cells per well of a 12-well plate pre-coated with fibronectin. Seven days post-plating triplicate wells were transfected with TransIT-TKO® (3-5 µl per well) and non-targeting control siRNA or GAPDH targeting siRNA (50nM per well). Seventy-two hours post-transfection, the amount of GAPDH mRNA was measured relative to 18s rRNA mRNA levels using qRT-PCR and then scaled to the expression level of the non-targeting control siRNA.  Error bars represent the standard error of the mean (SEM) of three independent complexes. See more information on stem cell applications.

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Delivery of Fluorescently-Labeled siRNA Using TransIT-TKO Transfection Reagent

Delivery of Fluorescently-Labeled siRNA Using TransIT-TKO® Transfection Reagent. HeLa (70% confluence) cells in 12-well plates were transfected with TransIT-TKO® Transfection Reagent (3 μl/well) and Label IT® siRNA Tracker Fluorescein-labeled siRNA duplexes (GREEN, 50 nM final concentration in the well). The cells were incubated 24 hours post-transfection then fixed and counterstained with TO-PRO®-3 (nuclei, BLUE) (Life Technologies) and Alexa Fluor® 546 Phalloidin (actin, RED) (Life Technologies). Confocal images were acquired on a Zeiss LSM 510 Confocal Microscope.

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High Efficiency Knockdown and Low Toxicity Using TransIT-TKO Reagent in HeLa Cells

High Efficiency Knockdown and Low Toxicity Using TransIT-TKO® Reagent in HeLa Cells. Firefly and sea pansy luciferase reporter vectors were co-transfected into HeLa cells using TransIT®-LT1 Reagent. Cells were incubated for at least 4 hours, split into 24-well plates, allowed to adhere and transfected with 25 nM of either a non-targeting siRNA or an anti-firefly luciferase siRNA using the indicated reagents with the volumes noted beneath each well. Luciferase expression, normalized to non-targeting siRNA control (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.

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Efficient Knockdown of Endogenous Genes in Primary Hepatocytes Using TransIT-TKO Reagent

Efficient Knockdown of Endogenous Genes in Primary Hepatocytes Using TransIT-TKO® Reagent. Primary mouse hepatocytes were transfected with the indicated siRNAs or a non-targeting control siRNA using the TransIT-TKO® Reagent. Twenty-four hours post-transfection, the amount of each mRNA was measured relative to GAPDH mRNA levels using qRT-PCR and then scaled to the expression level of the specific target mRNA in the cells alone (untreated) controls.

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Cell Line (Source) Endogenous Transcript Knockdown Efficiency
BNL CL.2 (mouse liver) MAPK1 80%
  MAPK3 83%
HeLa (human cervix) Lamin A/C 80%
  GAPDH 80%
Hepa1c1c7 (mouse liver) MAPK1 80%
  MAPK3 75%
  MEK1 75%
  PTEN 80%
HepG2 (human liver) MAPK1 80%
NIH 3T3-L1 MAPK1 70%
  MAPK3 70%
Secondary Human Astrocytes Lamin A/C 80%
Primary Mouse Hepatocytes ABC A1 70%
  Lamin A/C 81%

Knockdown of Endogenous Genes Using TransIT-TKO® Reagent. Various cells were transfected with siRNAs targeting the indicated genes using the TransIT-TKO® Reagent, and the knockdown percentage was determined using quantitative RT-PCR.

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Specifications

Storage Conditions:
All Configurations: Store at 4°C

Product Guarantee:
All Configurations: 1 year

Technical Product Literature


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Testimonials

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Review published in Select Science

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