產(chǎn)品簡(jiǎn)介：
PepMute?體外siRNA轉染試劑，是模擬病毒細胞穿膜肽配制的一種全新siRNA載體工具。在各種哺乳細胞中，在濃度只有1nM siRNA它卻能提供超過(guò)95%以上的沉默效率。運用我們專(zhuān)有的肽模擬合成技術(shù)，數以百計的病毒細胞穿膜肽被模擬、合成、篩選出來(lái)，并作為基因載體用在各種哺乳動(dòng)物的細胞系中（圖1）。PepMute?試劑作為一種非常有效的載體經(jīng)過(guò)鑒定和驗證，它能聚集和轉染短（小于100bp）、單鏈或雙鏈核苷酸如siRNA，miRNA 模擬物和單鏈DNA，且被廣泛用于哺乳動(dòng)物細胞系中。
Figure 1. A cartoon showing PepMute? siRNA Transfection Reagent was developed by PST
產(chǎn)品規格：
– PepMute? 試劑， 1.0 ml，轉染0.5~5 pmoles siRNA或miRNA模擬物，能在24孔板中足夠完成1333次反應。
– PepMute?轉染緩沖液（5X )，與PepMute? 試劑配合使用以獲得轉染效率最大化，8.0ml （5x )濃縮原液下能制成40 ml的工作液。
應用：
– siRNA, miRNA模擬物或 mRNA 轉染
– DNA/siRNA共轉染
– 單鏈DNA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)：
– 在最終濃度為1.0 nM siRNA也能保持非常出色的沉默效應。
– 使用在各種各樣的細胞系中，超過(guò)95%的有基因沉默效應。
– 單管反應，簡(jiǎn)單、標準的操作程序。
– 與血清和抗生素兼容。
– 適用于高通量篩選。
– 細胞毒性低。
– 價(jià)格實(shí)惠，物美價(jià)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.
Data Sheet & Protocol
– A Standard siRNA Transfection Protocol 
– A Standard siRNA/DNA Co-transfection Protocol
– A Reverse siRNA Transfection Protocol for HTS
To request a free trial sample, please Create An Account with us to enter your shipping address and email us at order@signagen.com
Testimonials:
I had tried your product pepMute transfection reagent on primary retinal neurons and was satisfied with the transfection efficiency. Will order!
– Vijai Krishnan Ph.D, Louisiana State University
I really appreciate you sending me a sample of PepMute siRNA reagent. I tested DNA/siRNA co-transfection using 293T cells and the results were completely satisfactory. I was able to get 95% knockdown of my target gene at 1.0 nM siRNA as well as expression of plasmid DNA using the recommended protocol. I will have to test and see if other cell lines are also as effective. I will definitely think about moving to use this reagent over Dharmafect or Oligofectamine.
– HARISH N. RAMANATHAN Ph.D., NIDDK, NIH
I tried PepMute reagent and I like it. It has so high efficiency and no cytotoxicity. I am going to use it. Thank for introducing it to our lab.
– Radmila Hrdlickova, Ph.D., UT Austin
Tried 3 different siRNAs with 100% silencing on A2780 cell. That is amazing! I already placed an order.
– Doris Benbrook, Ph.D., OUHSC
Yes, I certainly did use that sample of PepMute you generously sent us… and it worked so well on HepG2 cells that I have since ordered a full size and am using it exclusively for siRNA!? I compared to RNAiMAX and Roche’s X-tremeGENE products, but these products were not as effective as PepMute.? Thanks again for the sample.? It was fantastic!
– Matthew Jackson,? Ph.D., USDA
