TransIT-TKO¢ç Transfection Reagent
CAT# PRODUCT SIZE STORAGE ¼ÒºñÀÚ°¡ ÁÖ¹®
MIR2154[Mirus] TransIT-TKO¢ç Transfection Reagent0.4 ml-·Î±×ÀÎ
MIR2150[Mirus] TransIT-TKO¢ç Transfection Reagent1.5 ml-·Î±×ÀÎ
MIR2155[Mirus] TransIT-TKO¢ç Transfection Reagent5 X 1.5 ml-·Î±×ÀÎ
MIR2156[Mirus] TransIT-TKO¢ç Transfection Reagent10 X 1.5 ml-·Î±×ÀÎ
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TransIT-TKO¢ç Transfection Reagent

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

  • ±¤¹üÀ§ÇÑ ¹üÀ§ÀÇ siRNA¿Í miRNA Àü´Þ - ´Ù¾çÇÑ ¼¼Æ÷¿¡ siRNA¿Í miRNA¸¦ transfectionÇÒ ¼ö ÀÖÀ½.
  • µÎ °³ÀÇ ¼­·Î ´Ù¸¥ ½Ã¾à Á¶¼º - ÁÖ¾îÁø ¼¼Æ÷ÁÖ¿¡ È¿À²ÀûÀ¸·Î transfection ÇÒ ¼ö ÀÖ´Â ½Ã¾àÀ» Á¶¼º¿¡ µû¶ó µÎ °¡Áö Áß¿¡¼­ ¼±ÅÃÇÏ¿© »ç¿ë ÇÒ ¼ö ÀÖÀ½.
  • ³ôÀº knockdown È¿À² - ´ëºÎºÐÀÇ ¼¼Æ÷¿¡¼­ ÃÖÀûÀÇ gene silencing °á°ú¸¦ ¾òÀ½.
  • ³·Àº ¼¼Æ÷ µ¶¼º - ³·Àº µ¶¼ºÀ¸·Î ³ôÀº ¼¼Æ÷ »ýÁ¸À²À» À¯Áö½ÃÄÑ ½ÇÇèÀû ¿ÀÂ÷¸¦
    ÁÙÀÓ.
  • ´Ù¾çÇÑ ½ÇÇè¹æ¹ý - ´Ù¾çÇÑ protocol Àû¿ë °¡´É.
 
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TransIT-TKOⓇ°ú TransIT-siQUESTⓇ small RNA (siRNA and duplex miRNA) Transfection ½Ã¾àÀº ±¤¹üÀ§ÇÏ°Ô »ç¿ëµÇ´Â Á¦Ç°À̸ç ÃÖ¼ÒÀÇ ¼¼Æ÷ µ¶¼ºÀ» °¡Áö°í ÀÖ½À´Ï´Ù. ÀÌ µÎ ½Ã¾àÀº »ç¿ëÀÚ°¡ ÀÚ½ÅÀÇ Æ¯Á¤ÇÑ ¼¼Æ÷ÁÖ¿¡ ÀûÇÕÇÑ ÃÖ»óÀÇ transfection ½Ã¾àÀ» Á¶¼º¿¡ µû¶ó ¼±Åà ÇÏ¿© »ç¿ëÇÒ ¼ö ÀÖ½À´Ï´Ù.
 
Figure 1. High Efficiency Endogenous Knockdown in iCell¢ç Cardiomyocytes. The TransIT-TKO¢ç 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¢ç (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¢ç Transfection Reagent. HeLa (70% confluence) cells in 12-well plates were transfected with TransIT-TKO¢ç Transfection Reagent (3 ¥ìl/well) and Label IT¢ç 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¢ç-3 (nuclei, BLUE) (Life Technologies) and Alexa Fluor¢ç 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¢ç Reagent in HeLa cells. Firefly and sea pansy luciferase reporter vectors were co-transfected into HeLa cells using TransIT¢ç-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¢ç Reagent. Primary mouse hepatocytes were transfected with the indicated siRNAs or a non-targeting control siRNA using the TransIT-TKO¢ç 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¢ç Reagent. Various cells were transfected with siRNAs targeting the indicated genes using the TransIT-TKO¢ç Reagent, and the knockdown percentage was determined using quantitative RT-PCR.
Storage Conditions: Store at 4¡ÆC
Product Guarantee: 1 year
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