Fluc mRNA with N1-Me-pUTP (5’CAP) |
| Katalog-Nr.GM10005 |
Fluc mRNA with N1-Me-pUTP (5'CAP) is labeled luciferase mRNA produced by in vitro transcription, has a Cap 1 cap structure and a poly(A) tail, and contains N1-Me-pUTP modification.
Products are for research use only. Not for human use. We do not sell to patients.
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
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Luciferase(Fluc) can detect gene expression extremely sensitively and efficiently, so it is often used as a bioluminescent reporter gene for gene regulation and functional studies. Fluc 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. Firefly luciferase protein catalyzes intracellular luciferin or fatty aldehyde to produce autofluorescence, producing chemiluminescence at a wavelength of about 550-570nm[1].
Luciferase can be used to detect promoter activity or dual fluorescent molecule complementation experiments. Firefly luciferase and Renilla luciferase can respectively catalyze their respective substrates to emit fluorescence of different colors. The two light absorption wavelengths are different and the detection does not interfere with each other. Therefore, they can be used as dual luciferases in the same chemical reaction system. Reporter gene systems are used simultaneously[2].
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 increases the stability and translation efficiency of the mRNA[3]. N1-Me-pUTP is a methyl modification of the naturally occurring pseudouridine pUTP, which is catalyzed by the N1-specific pseudouridine methyltransferase Nepl that exists in archaea and eukaryotes[4]. This product uses N1-Me-pUTP instead of UTP, which effectively enhances RNA stability and reduces anti-RNA immune responses[5].
References:
[1]. JoÃo M M LeitÃo, Joaquim C G Esteves da Silva. Firefly luciferase inhibition. 2010 Oct 5;101(1):1-8. doi: 10.1016/j.jphotobiol.2010.06.015. Epub 2010 Jul 3.
[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]. 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.
[4]. 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.
[5]. 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.
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| Buffer | Storage | -40°C or below | |
| General tips | Please dissolve it on ice and be careful to prevent RNase contamination and degradation. Avoid repeated freezing and thawing as much as possible. Do not vortex. For first use, gently centrifuge and divide into portions for individual use. Use RNase-free reagents and consumables, use appropriate RNase-free techniques, and do not add to the serum-containing medium until mixed with the transfection reagent. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
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Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)
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