OVA mRNA with N1-Me-pUTP (5'CAP)
目录号 : GM10012OVA mRNA with N1-Me-pUTP(5'CAP)是通过体外转录产生的卵白蛋白mRNA,具有Cap 1帽结构和poly(A)尾,并含有N1-Me-pUTP修饰。
Sample solution is provided at 25 µL, 10mM.
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Ovalbumin (OVA) is a member of the chromoprotein superfamily and the main protein component in egg white. OVA is a glycoprotein with a molecular weight of approximately 45,000 daltons, which can induce moderate immunity in the body and is a commonly used antigen in immune and biochemical research. Ovalbumin is often used as an immunogen for immune experiments, such as the establishment of animal models such as altitude sickness and asthma. OVA mRNA can express proteins directly in the cytoplasm without relying on a promoter. The protein expression speed is faster than transfected DNA. The protein expression level is directly related to the transfection amount of mRNA, and there is no risk of gene integration.
OVA mRNA with N1-Me-pUTP is produced through in vitro transcription. By simulating the mRNA processing process in eukaryotes, the product has a Cap 1 cap structure at the 5' end and a poly(A) tail at the 3' end, which can reduce the immune response of the host cell and increase the amount of mRNA. stability and translation efficiency [1]. N1-methyl-pseudouridine (1-methylpseudouridine, m1ψ) is a methyl modification of naturally occurring pseudouridine, formed by N1-specific pseudouridine A that exists in archaea and eukaryotes It is catalyzed by the base transferase Nepl[2]. N1-methyl-pseudouridine is a substitute for uridine (U), which can effectively enhance RNA stability while reducing anti-RNA immune response [3].
References:
[1]. Jemielity J, Fowler T, Zuberek J, et al. Novel "anti-reverse" cap analogs with superior translational properties. RNA. 2003;9(9):1108-1122.
[2]. Callum J C Parr, et al.N 1-Methylpseudouridine substitution enhances the performance of synthetic mRNA switches in cells. 2020 Apr 6;48(6):e35. doi: 10.1093/nar/gkaa070.
[3]. Pedro Morais, Hironori Adachi, Yi-Tao Yu.The Critical Contribution of Pseudouridine to mRNA COVID-19 Vaccines. 2021 Nov 4;9:789427. doi: 10.3389/fcell.2021.789427.
卵白蛋白(Ovalbumin,OVA)是色蛋白超家族的成员,也是蛋清中的主要蛋白成分。OVA是一种分子量约45000道尔顿的糖蛋白,能够引起机体适度的免疫性,是免疫和生化研究的常用抗原。卵白蛋白常被用作免疫原进行免疫实验,如高原反应性、哮喘等动物模型建立。OVA mRNA能够在不依赖于启动子的情况下直接在细胞质中表达蛋白,蛋白表达速度比转染DNA更快,蛋白表达量与mRNA的转染量直接相关,并且没有基因整合的风险。
OVA mRNA with N1-Me-pUTP通过体外转录产生,通过模拟真核生物中mRNA加工过程,该产品的5'端具有Cap 1帽结构,3'端具有poly(A)尾,可以减少宿主细胞的免疫反应并增加mRNA的稳定性和翻译效率[1]。N1-甲基-假尿苷(1-甲基假尿苷,m1ψ)是天然存在的假尿苷的甲基修饰物,由存在于古细菌和真核生物中的N1特异性假尿苷甲基转移酶Nepl催化生成[2]。N1-甲基-假尿苷是尿苷(U)的替代物,能够有效增强RNA稳定性,同时降低抗RNA免疫应答[3]。
mRNA Length | 1438 nucleotides | ||
Concentration | 1mg/mL | ||
Buffer | 1 mM Sodium Citrate, pH 6.4 | 储存条件 | -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,才可加入合有血清的培养基中。 |
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Shipping Condition | 评估样品解决方案:配备蓝冰进行发货。所有其他可用尺寸:配备RT,或根据请求配备蓝冰。 |
第一步:请输入基本实验信息(考虑到实验过程中的损耗,建议多配一只动物的药量) | ||||||||||
给药剂量 | mg/kg | 动物平均体重 | g | 每只动物给药体积 | ul | 动物数量 | 只 | |||
第二步:请输入动物体内配方组成(配方适用于不溶于水的药物;不同批次药物配方比例不同,请联系GLPBIO为您提供正确的澄清溶液配方) | ||||||||||
% DMSO % % Tween 80 % saline | ||||||||||
计算重置 |
计算结果:
工作液浓度: mg/ml;
DMSO母液配制方法: mg 药物溶于 μL DMSO溶液(母液浓度 mg/mL,
体内配方配制方法:取 μL DMSO母液,加入 μL PEG300,混匀澄清后加入μL Tween 80,混匀澄清后加入 μL saline,混匀澄清。
1. 首先保证母液是澄清的;
2.
一定要按照顺序依次将溶剂加入,进行下一步操作之前必须保证上一步操作得到的是澄清的溶液,可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。