產(chǎn)品簡(jiǎn)介：
GenMute? siRNA試劑是一種新型的可生物降解的非脂質(zhì)體的siRNA / miRNA和DNA的載體工具。運用我們專(zhuān)有的PH依賴(lài)構象變化技術(shù)，這款可生物降解的轉染試劑在它的側鏈上添加了預篩選出來(lái)的疏水集團，使得GenMute?試劑成為市場(chǎng)上最萬(wàn)能的最強大的基因載體工具。GenMute?試劑，已被證實(shí)能有效的，可重復性轉染單siRNA片段和miRNA模擬物或共轉染DNA/siRNA片段或DNA/miRNA片段到各種哺乳動(dòng)物細胞中。
產(chǎn)品規格：
– GenMute?轉染試劑，1.0ml，足夠在24孔板中轉染5.0 pmoles的siRNA或miRNA模擬物達到2000個(gè)反應。
– GenMute? 轉染試劑緩沖液（5X），與PGenMute? 試劑配合使用以獲得轉染效率最大化，8.0ml（5X）濃縮液就可以制成40ml的工作液體。
應用：
– 對各種哺乳動(dòng)物細胞的單siRNA 轉染或者 DNA/siRNA 共轉染。
– 對各種哺乳動(dòng)物細胞的單 miRNA， miRNA模擬物 或者 miRNA抑制劑的轉染或者 DNA/miRNA 或DNA/miRNA模擬物的共轉染。
儲存條件：
40C儲存。若儲存合適，產(chǎn)品的穩定性能保持12個(gè)月以上。
特點(diǎn)：
– 在只有 1.0 nM siRNA / miRNA模擬物的低濃度中，也能達到非常好的沉默效應。
– 在細胞內，能將非特異性和非靶目標的影響降到最低。
– 單管反應。
– 運用于廣泛的哺乳動(dòng)物細胞系。
– 細胞毒性低。
– 物美價(jià)廉，非常實(shí)惠。
Comparisons of Silencing Efficacy of GenMute? siRNA & DNA Transfection Reagent with Brand Name Products
Knockdown efficacy comparison of GenMute? Transfection Reagent (upper panel) vs. Dharmafect? 4 siRNA Transfection Reagent (middle panel) and Lipofectamine? 2000 (lower panel) on HEK293 cells. siRNA targeting renilla luciferase at different final concentrations ranging from 0.5 to 20 nM was co-transfected with renilla luciferase gene (0.5 μg of pRL-CMV DNA per well) by the above three transfection reagents per manufacturers’ protocols into HEK293 cells growing on a 24-well plate. Renilla luciferase activity was determined 24h after post co-transfection with renilla luciferase determination system (Promega). The luminescence was measured from 5.0 μl of lysate during 10s integration with a luminometer (Beckman Coulter LD 400). Luciferase activity was expressed as light units integrated over 10s (RLU) and normalized per mg of cell protein by using the BCA assay. The errors bars represent standard deviation derived from triplicate experiments. Luciferase-silencing efficiency was calculated relative to untreated cells. While GenMute? and Dharmafect? 4 reagents delivered significant gene silencing from 1.0 nM of renilla luciferase siRNA, lipofectamine? 2000 gave good knockdown only after 30 nM (data not shown).
Excellent silencing of endogenously expressed KIF11 (also known as Eg5) in HEK293 cells with 0.5 μl of GenMute? reagent and 5.0 pmol Eg5 siRNA per well of 24-well plate. KIF11 (also known as Eg5) encodes a motor protein that belongs to the kinesin-like protein family involved in chromosome positioning and bipolar spindle formation during cell mitosis. A reduction in KIF11 levels causes mitotic arrest. GenMute? reagent effectively delivers Eg5 siRNA (final 10 nM, right panel) to HEK293 cells at only 0.5 μl per well of 24-well plate vs. negative control (final 10 nM. left panel). Compared with negative control (left panel), phenotype of “rounded-up” 293 cells were visualized 24 hours post transfection (right panel) with a Nikon microscope.
GenMute? Transfection Reagent knocked down endogenous lamin A/C gene expression in Hela cells. A siRNA targeting lamin A/C gene (right panel) and a sham siRNA (left panel) were introduced into Hela cells (final 1.0 nM) by GenMute? Transfection Reagent. Lamin A/C gene silencing was monitored 24h post transfection by immunofluorescence. Lamin A/C was probed with a mouse monoclonal lamin A/C specific antibody followed by addition of FITC-conjugated anti-mouse antibodies. Quantitative analysis showed that lamin siRNA at 1.0 nM delivered by GenMute? Transfection Reagent knocked down 94% endogenously expressed lamin A/C in Hela cells.
Data Sheet & Protocol
– A Protocol for siRNA Transfection and DNA/siRNA Co-Tranafection 
– A Protocol for miRNA Mimics Transfection and DNA/miRNA Co-Transfection
– A Protocol for DNA Transfection to Mammalian Cells
