PepMute™体外siRNA转染试剂

价格: ¥2,080.00

购买数量:

  • 目录号: SL100566
  • 包装: 1.0 mL

请选择:



产品简
PepMute™体外siRNA转染试剂,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 siRNAmiRNA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。储存合适,产品的稳定性能保持12个月以上

特点
-
在最浓度为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。

PepMute™ siRNA转染试剂和领先品牌产品沉默效应的比较

2: 在24孔板中,每孔加1.0 µl PepMute™试剂和0.5 pmol EG5 siRNA, 内源性表达基因KIF11(也称EG5)在HEK293表现出出色的沉默效应。KIF11 (也称EG5)编码了一个启动蛋白,这个启动蛋白属于驱动蛋白家族,参与染色体定位和细胞有丝分裂时两端的纺锤体的形成。KIF11水平降低会阻碍有丝分裂。 PepMute™试剂能有效的将EG5 siRNA (终浓度 1.0 nM)转染到HEK293细胞中,转染48小时后,参照阴性对照(终浓度是1.0 nM+假EG5 siRNA),在HEK293细胞上能导致大于 80%的圆顶表型。然而其他主要的siRNA转染试剂,在HEK293细胞上,Lipofectamine™ RNAiMAX (RNAiMAX, 1.0 nM EG5 siRNA) / INTERFERin (1.0 nM EG5 siRNA) / Dharmafect (10.0 nM EG5 siRNA)和jetPRIME (20 nM EG5 siRNA)只能得到平均37%, 23%, 53% 和48%圆顶表型。转染48小时后,通过尼康荧光显微镜观察293细胞上的圆顶表型,可视化的图片(上图)和量化的图片(下图)。



图3: PepMute™转染试剂(上图)和Dharmafect 4 (中图)以及Lipofectamine™ RNAiMAX (RNAiMAX,下图) siRNA转染试剂在A549细胞上沉默效率的对比。siRNA抑制的海肾荧光素酶在不同的最终浓度范围(0.5~20nM)与海肾荧光素酶基因(0.5 µg pRL-CMV DNA/孔,24孔板)通过上述的三种转染试剂按照生产制造商的科学实验步骤共转染入 A549细胞。共转染36小时后,通过海肾荧光酶测定系统(Promega)检测海肾荧光酶的活性。通过5 µl 裂解液10s的裂解整合,运用光度计(贝克曼库尔特LD 400)测量荧光。荧光酶活性的表达检测通过超过10s相对光单位(RLU)检测和采用BCA检测标准化的每毫克细胞蛋白含量。误差来自于重复的三次实验的标准偏差。计算未处理的细胞荧光素酶沉默效应。PepMute™和 Dharmafect™ 4试剂, 1.0 nM海肾荧光素酶siRNA 能产生显著地基因沉默效应Lipofectamine™ RNAiMAX只有在浓度为20nM时,具有良好的基因敲减作用。然而在( 0.5~1.0 nM)的低浓度下,能否提高基因的表达还有待于观察。



图4: PepMute™转染试剂反向分别转染能5.0和1.0 nM GFP siRN到MCF7细胞(上图)和U2OS细胞(下图)中,能得到大大降低稳定的GFP表达。绿色荧光蛋白(GFP)能稳定的在MCF7和U2OS细胞中表达。通过PepMute™转染试剂将终浓度为5.0nM   和1.0 nM的siRNA抑制的GFP基因(右图)和假siRNA (左图)被分别转导入MCF7和U2OS细胞中。 转染48小时后通过尼康荧光显微镜观察GFP基因沉默。定量分析表明,通过 PepMute™ siRNA转染试剂将5.0nM和1.0 nM siRNA抑制的GFP基因分别反转染到 MCF7和U2OS细胞中能达到90%和95%的稳定表达。



转染操作步骤:
- 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 [email protected]


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

 

 




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