mRNA Cap 2'-O-Methyltransferase, GMP Grade (50U/μl)

- Overview
  - With S-adenosylmethionine (SAM) as the methyl group base donor, mRNA Cap 2´-O-Methyltransferase can add a methyl group to the 2´-O of the first nucleotide of the Cap0 structure, [m7GpppN1(pN)x-OH(3')], next to the RNA 5´ end. Methylation of capped mRNA from Cap 0 to Cap1 naturally exists in eukaryocyte, where the Cap1 structure shows an important effect for translation, thus for expression as well. This product doesn’t show a reactivity to RNA substrate other than RNA with Cap0 structure.
    This product is from a large scale GMP leveled recombinant Cap 2’-O-Methyltransferase production via E. coli expression. Applying pharmaceutical leveled adjuvant and material for production, strictly controlling host protein residues, nucleic acid residues and other impurities, we guarantee manufacture and quality control practice complying to GMP regulation, as well as all the materials traceable.
    This product has completed the DMF record of FDA and passed the HALAL certification.
    
    Please contact us at for specific academic pricing.
    
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    FDA DMF Filed Enzymes
- Properties
  - Storage
    
    At -20±5 °C.
    
    Note
    
    SAM is unstable at pH 7–8, 37 °C and needs to be freshly prepared before the reaction starts. The amount of SAM can be calculated in advance, and the aliquoted 32 mM stock solution is diluted into a 2 mM working solution before the reaction starts. To avoid SAM degradation, the working solution needs to be kept on ice.
    Before starting reaction, the RNA applied for capping should be purified and dissolved in nuclease-free water. The solution for use must NOT contain any EDTA or salt.
    RNA transcript with 5’ end secondary structure can be an obstacle for capping. Heating the RNA sample can remove complex structure of the transcript. We may extend heating time to 10 minutes and extend the capping reaction time to 60 minutes.
    Capping from Cap0 to Cap1 naturally exists in eukaryotic cells, which enhance translation efficiency.
    Vaccinia Capping Enzyme (GMP-M062) can co-work with 2’-O-Methyltransferase (GMP-M072). When using new cell lines or translation systems, it is recommended to compare the translation efficiency and immunogenicity of Cap 0 and Cap 1-mRNA to determine the optimal Cap structure.
    When adding 0.5 µl of RNase inhibitor (GMP-E125, Novoprotein) to the reaction system (shown below in “Manual” part), please deduct equal volume of RNase-free water.
    When used for RNA 5´ -end labeling, GTP should be diluted to 1-3 times of the mol concentration of mRNA.
    When used for the capped RNA 3’-poly (A) tailing, we may apply E. coli Poly(A) Polymerase, GMP Grade (GMP-M012, Novoprotein).
    The sample needs to be purified before transfection.
    Do NOT store the product at -70°C or an even lower temperature.
    In case of white precipitation in reaction buffer (shown below in “Manual” part), incubate it at 37 ° C for 5 mins, blend it, totally dissolving precipitation.
    
    Expression system
    
    E. coli
    
    Endotoxin
    
    ＜5EU/ml
    
    Activity
    
    50KU/ml-52.5KU/ml
    
    * For Research or Manufacturing Use Only, Not for Use in Diagnostic or Therapeutic Procedures.

Datasheet

mRNA Cap 2'-O-Methyltransferase, GMP Grade (50U/μl)

mRNA Cap 2'-O-Methyltransferase, GMP Grade (50U/μl)

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