ARCA EGFP mRNA
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 EGFP mRNA是一种报告(reporter)mRNA,可以用作对照,通过各种基于荧光的方法研究哺乳动物细胞中的转染和表达。
将ARCA EGFP mRNA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。
该产品已经是5’端加帽和3’端加poly(A)尾的mRNA,可以模拟真核生物中加工成熟的mRNA。使用ARCA(抗反向帽类似物,货号B8175)以共转录的方式对mRNA进行了加帽,使其具有Cap 0结构,增加了mRNA的稳定性和翻译效率[1,2]。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.
- 1. Yan Liang, Jingge Zhang, et al. "Biomimetic Mineralized CRISPR/Cas RNA Nanoparticles for Efficient Tumor-Specific Multiplex Gene Editing." ACS Nano. 2023 Aug 8;17(15):15025-15043. PMID: 37481734
- 2. Qiming Yin, Xiang Song, et al. "Incorporation of glycyrrhizic acid and polyene phosphatidylcholine in lipid nanoparticles ameliorates acute liver injury via delivering p65 siRNA." Nanomedicine. 2022 Dec 27;48:102649. PMID: 36584740
Identity & Purity
- 批次:
-
Agarose Gel Mobility: Pass.
- QC
- MSDS (Material Safety Data Sheet)
- Datasheet
Concentration ± 6%: Pass.
相关生物数据
mRNA Length | 996 nucleotides | ||
Concentration | 1 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。 |
细胞系 | HEK293T |
细胞培养板 | 48孔板 |
铺板细胞量 | 5w左右 |
转染前细胞密度 | 细胞融合度50%-60% |
转染条件 | 培养17h后在无血清无抗生素的培养基中进行转染(1μg mRNA+2μL lipofectamine 2000),4h后更改为完全培养基恢复至正常生长环境,24h后进行荧光拍摄 |
RNA转染量和转染试剂 | 1μg mRNA与2μL lipofectamine 2000按照说明书制备 |
转染效果 | 转染效率90%以上 |