產(chǎn)品介紹：
PepMute? Plus體外siRNA和DNA轉染試劑是PepMute?轉染試劑(目錄號：SL100566)的升級版。 PepMute? Plus體外siRNA和DNA轉染試劑是在PepMute?轉染試劑的基礎上經(jīng)過(guò)其骨架添加輸水性氨基酸所得到的?；谖覀儶毺氐?/span>pH依賴(lài)的分子構象轉化技術(shù)，我們在36個(gè)氨基酸組成的骨架多肽基礎上經(jīng)過(guò)添加輸水性的氨基酸并經(jīng)過(guò)反復篩選得到，這種化學(xué)修飾賦予PepMute? Plus試劑一種獨特性質(zhì)，即在形成轉染復合物時(shí)該試劑獲得了自我裝配的特性。這一特性使得該試劑成為一個(gè)最為高效的轉染試劑，實(shí)驗證明，該試劑對siRNA有及其高效的轉染效率，即使在siRNA終濃度為1.0 nM時(shí)，也可以獲得95%基因沉默效率。?PepMute? Plus試劑已經(jīng)被證明可有效地對多種哺乳動(dòng)物細胞進(jìn)行有效的siRNA和DNA轉染以及和DNA/siRNA共轉染。
Figure 1. A cartoon showing PepMute? Plus siRNA Transfection Reagent was developed
Size & content:
– PepMute? Plus Transfection Reagent, 1.0 ml, sufficient for ~1000 reactions based on transfecting 2.5 pmols siRNA in 24-well plate
– PepMute? Plus Transfection Buffer(5x ), formulated for maximal transfection efficiency, 8.0 ml
Storage:
Store at 4 °C and never freeze. If stored properly, the product is stable for 12 months or longer.
Advantages:
– Effective knockdown (95%) with only 0.5 pmol siRNA (1.0 nM siRNA in 24-well plate)
– Formulated for hard-to-transfect mammalian cells
– One tube reaction and easy transfection protocol
– Best for broad spectrum especially hard-to-transfect mammalian cells
– Very low cytotoxicity
Comparisons of Knockdown Efficacy of PepMute? Plus siRNA & DNA Transfection Reagent with Brand Name Products
Figure 2. Knockdown efficacy comparison of PepMute? Plus 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 PepMute? Plus 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).
Figure 3. PepMute? Plus Transfection Reagent knocked down GFP gene expression in HepG2 cells. GFP cDNA, pEGFP-N3, was co-transfect with a siRNA targeting GFP gene (final 5.0 nM, right panel) and a sham siRNA (final 5.0 nM, left panel) into HepG2 cells by PepMute? Plus Transfection Reagent. GFP fluorescence was visualized 24h post transfection with a Nikon T2000 fluorescence micorscopy. Quantitative analysis showed that GFP siRNA at 5.0 nM delivered by PepMute? Plus Transfection Reagent knocked down 96% co-transfected GFP expression in HepG2 cells.
轉染操作步驟?
– A Protocol for DNA/siRNA Co-Tranafection to Mammalian Cells 
– A Protocol for siRNA Transfection to Mammalian Cells
– A Protocol for DNA Transfection to Mammalian Cells
