TransIT-TKO Transfection Reagent
CAT# PRODUCT SIZE STORAGE Һڰ ֹ
MIR2154[Mirus] TransIT-TKO Transfection Reagent0.4 ml-α
MIR2150[Mirus] TransIT-TKO Transfection Reagent1.5 ml-α
MIR2155[Mirus] TransIT-TKO Transfection Reagent5 X 1.5 ml-α
MIR2156[Mirus] TransIT-TKO Transfection Reagent10 X 1.5 ml-α
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TransIT-TKO Transfection Reagent

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

  • siRNA miRNA - پ siRNA miRNA transfection .
  • ٸ þ - ־ ֿ ȿ transfection ִ þ ߿ Ͽ .
  • knockdown ȿ - κ gene silencing .
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  • پ - پ protocol .
 
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TransIT-TKOⓇ TransIT-siQUESTⓇ small RNA (siRNA and duplex miRNA) Transfection þ ϰ Ǵ ǰ̸ ּ ֽϴ. þ ڰ ڽ Ư ֿ ֻ transfection þ Ͽ ֽϴ.
 
Figure 1. 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.
Figure 2. 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 l/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.
Figure 3. 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.
Figure 4. 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.
 
Cell Line (Source)    Endogenous Transcript  Knockdown Efficiency 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%
Figure 5. 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.
Storage Conditions: Store at 4C
Product Guarantee: 1 year
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Mirus TKO transfection reagent
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