ARCA Cy3 EGFP mRNA (5-moUTP)
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
ARCA Cy3 EGFP mRNA (5-moUTP) 可用于分析mRNA的递送和翻译效率。将ARCA Cy3 EGFP mRNA (5-moUTP)转染到细胞后,细胞可表达EGFP(增强型绿色荧光蛋白)。EGFP蛋白是一种常见的报告分子,激发时会发出强烈且明亮的绿色荧光,其最大激发/发射光波长分别为488 nm /509 nm。该mRNA带有荧光染料Cyanine 3(简称Cy3)标记,用一定波长的光激发时可以直接可视化EGFP mRNA,Cy3的最大激发和发射波长分别为550 nm和570 nm。因此Cy3 EGFP mRNA是独立于翻译确定mRNA递送和定位的理想分子。
该产品已经是5’端加帽,3’端加poly(A)尾,以及经过5-moUTP和Cyanine 3-UTP修饰的mRNA。Cyanine 3-UTP与5-moUTP的添加比例是1:3。使用ARCA(抗反向帽类似物,货号B8175)以共转录的方式对mRNA进行加帽,形成了Cap 0结构,增加了mRNA的稳定性和翻译效率[1,2]。修饰核苷酸5-moUTP(也称为5-methoxy-UTP,货号B8061)的添加可以抑制RNA介导的先天免疫激活。Poly(A)尾的添加使mRNA更加稳定并提高mRNA的翻译起始效率[3]。
mRNA转染的优点:
· 无需核吸收——蛋白直接在细胞质中表达
· 其蛋白质表达比DNA转染更快
· 以完全不依赖启动子的方式表达蛋白质
· 没有基因组整合的风险
· 非常适合转染生长缓慢或不分裂的细胞
· 蛋白表达与mRNA的量直接相关
· 瞬时转染:蛋白质的表达在有限的时间内持续,避免了与积累有关的毒性
References:
[1] Stepinski J, Waddell C, Stolarski R, et al. Synthesis and properties of mRNAs containing the novel "anti-reverse" cap analogs 7-methyl(3'-O-methyl)GpppG and 7-methyl (3'-deoxy)GpppG. RNA. 2001;7(10):1486–1495.
[2] Jemielity J, Fowler T, Zuberek J, et al. Novel "anti-reverse" cap analogs with superior translational properties. RNA. 2003;9(9):1108–1122.
[3] Gallie DR, Tanguay R. Poly(A) binds to initiation factors and increases cap-dependent translation in vitro. J Biol Chem. 1994;269(25):17166–17173.
mRNA Length | 996 nucleotides | ||
Concentration | 1.0 mg/mL | ||
Buffer | 1 mM Sodium Citrate, pH 6.4 | Storage | -40°C or below |
General tips | 请将其于冰上溶解,并小心防止RNase污染降解。尽可能避免反复冻融。 不要涡旋震荡。首次使用时,将其轻柔离心并分成几份,可供单独使用。使用不含RNase的试剂和耗材,使用适当的无RNase技术。直至与转染试剂混合,才可加入含有血清的培养基中。 | ||
Shipping Condition | 试用装:干冰运输。 |