TransIT-siQUEST¢ç Transfection ReagentA high efficiency, low toxicity, siRNA transfection reagent for mammalian cells
Ư¡
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 Knockdown and Low Toxicity Using TransIT-siQUEST¢ç Reagent in CHO Cells Stably Expressing Firefly Luciferase. CHO-luc cells were grown in 24 wells plates and transfected with 25 nM of either a non-targeting siRNA or a 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 percent 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 2. Inhibition of PPAR-alpha Expression in Primary Mouse Hepatocytes Using the TransIT-siQUEST¢ç Reagent. Primary mouse hepatocytes were transfected with an anti-PPAR-alpha siRNA or a non-targeting control siRNA using the TransIT-siQUEST¢ç Reagent. Twenty-four hours post-transfection, the amount of PPAR-alpha 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 Opti-MEM¢ç media only (untreated) control.
Figure 3. Delivery of Fluorescently-Labeled siRNA using TransIT-siQUEST¢ç Transfection Reagent. HeLa (70% confluence) cells in 12-well plates were transfected with TransIT-siQUEST¢ç Transfection Reagent (3 ¥ìl/well) and Label IT¢ç siRNA Tracker Cy-3-labeled siRNA duplexes (RED, 25 nM final concentration in the well). The cells were incubated 24 hours post-transfection then fixed and counterstained with counterstained with Alexa Fluor¢ç 488 Phalloidin (GREEN) (Life Technologies). Confocal images were acquired on a Zeiss LSM 510 Confocal Microscope.
Figure 4. Efficient Target Gene Knockdown in Selected Cell Lines Using TransIT-siQUEST¢ç Reagent. Reporter plasmids expressing both firefly and sea pansy luciferase were co-transfected into the indicated cell lines using TransIT¢ç Plasmid Transfection Reagents. Targeted knockdown was achieved by transfection of an anti-firefly luciferase siRNA using the TransIT-siQUEST¢ç Reagent. Twenty-four hours later, firefly luciferase expression was normalized to sea pansy luciferase expression and compared to the reagent alone control.
Storage Conditions: Store at 4¡ÆC
Product Guarantee: 1 year ml033_transit_siquest_transfection_reagent.pdf (423,184kb) |
»ùÇà ½ÅûÇϱâ
»ç¿ëÈıâ ÀÛ¼º
|