PepMute™体外siRNA转染试剂 进口转染试剂

美国Signagen转染试剂
货号SL100566

PepMute™体外siRNA转染试剂

产品简介：
PepMute™体外siRNA转染试剂，是模拟病毒细胞穿膜肽配制的一种全新siRNA载体工具。在各种哺乳细胞中，在浓度只有1nM siRNA它却能提供超过95%以上的沉默效率。运用我们专有的肽模拟合成技术，数以百计的病毒细胞穿膜肽被模拟、合成、筛选出来，并作为基因载体用在各种哺乳动物的细胞系中（图1）。PepMute™试剂作为一种非常有效的载体经过鉴定和验证，它能聚集和转染短（小于100bp）、单链或双链核苷酸如siRNA，miRNA 模拟物和单链DNA，且被广泛用于哺乳动物细胞系中。



Figure 1. A cartoon showing PepMute™ siRNA Transfection Reagent was developed by PST

产品规格：
- PepMute™ 试剂， 1.0 ml，转染0.5~5 pmoles siRNA或miRNA模拟物，能在24孔板中足够完成1333次反应。
- PepMute™转染缓冲液（5X )，与PepMute™ 试剂配合使用以获得转染效率最大化，8.0ml （5x )浓缩原液下能制成40 ml的工作液。

应用：
- siRNA, miRNA模拟物或 mRNA 转染
- DNA/siRNA共转染
- 单链DNA转染

储存条件：
4⁰C储存。若储存合适，产品的稳定性能保持12个月以上。

特点：
- 在最终浓度为1.0 nM siRNA也能保持非常出色的沉默效应。
- 使用在各种各样的细胞系中，超过95%的有基因沉默效应。
- 单管反应，简单、标准的操作程序。
- 与血清和抗生素兼容。
- 适用于高通量筛选。
- 细胞毒性低。
- 价格实惠，物美价廉。

Silencing Efficacy Comparison between PepMute™ siRNA Transfection Reagent and Leading Products

Figure 2. Excellent silencing of endogenously expressed KIF11 (also known as EG5) in HEK293 cells with 1.0 µl of PepMute™ reagent and 0.5 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. PepMute™ reagent effectively delivers EG5 siRNA (final 1.0 nM) to HEK293 cells, leading to more than 80% of "round-up" phenotype of HEK293 cells 48h post transfection over negative control (final 1.0 nM with sham EG5 siRNA) while leading siRNA transfectin reagents, Lipofectamine™ RNAiMAX (RNAiMAX, 1.0 nM EG5 siRNA) / INTERFERin (1.0 nM EG5 siRNA) / Dharmafect (10.0 nM EG5 siRNA) and jetPRIME (20 nM EG5 siRNA) give average 37%, 23%, 53% and 48% ball-shaped phenotype respectively on HEK293 cells. The phenotype of "rounded-up" 293 cells were visualized (upper panel) and quantified (lower panel) 48h post transfection with a Nikon microscope.


Figure 3. Silencing efficiency comparison of PepMute™ Transfection Reagent (upper panel) with Dharmafect 4 (middle panel) and Lipofectamine™ RNAiMAX (RNAiMAX, lower panel) siRNA Transfection Reagents on A549 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 A549 cells growing on a 24-well plate. Renilla luciferase activity was determined 36h 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™ and Dharmafect™ 4 reagents delivered significant gene silencing from 1.0 nM of renilla luciferase siRNA, Lipofectamine™ RNAiMAX gave good knockdown only after 20 nM while enhanced gene expression at low concentration of siRNA (0.5 and 1.0 nM respectively) was observed.



Figure 4. PepMute™ Transfection Reagent knocked down stable GFP expression in MCF7 cell (upper panel) and U2OS cell (lower panel) by reversely transfecting 5.0 and 1.0 nM GFP siRNA respectively. Green fluorescence protein (GFP) was stably expressed in MCF7 and U2OS cells. siRNA targeting GFP gene (right panel) and a sham siRNA (left panel) were introduced into MCF7 and U2OS cells with final 5.0 and 1.0 nM respectively by reverse transfection with PepMute™ Transfection Reagent. GFP gene silencing was monitored 48h post transfection by a Nikon fluorescence microscope. Quantitative analysis showed that GFP siRNA at 5.0 and 1.0 nM delivered by PepMute™ siRNA Transfection Reagent knocked down 90% and 95% stably expressed GFP in MCF7 and U2OS cells respectively.