Vaccinia Capping Enzyme (mRNA 5' Capping, Cap 0/Cap 1)
Recombinant vaccinia capping enzyme (D1-D12) with three-in-one activity for enzymatic mRNA 5' capping — ~100% Cap 0 in 1 hour, upgradeable to Cap 1. Low endotoxin, low host residuals, RUO.
Catalog no. VCE-500U/10KU/100KU/500KU/1000KU · Ships with certificate of analysis
For Research Use OnlyOverview
At a Glance
Recombinant vaccinia capping enzyme (D1 + D12 subunits) combining three catalytic activities in one enzyme: RNA triphosphatase, guanylyltransferase and guanine methyltransferase. It adds the native eukaryotic 7-methylguanosine cap (m7GpppN — Cap 0) to the 5' end of RNA, dramatically improving mRNA stability, nuclear export and translation efficiency. Co-reacted with a 2'-O-methyltransferase and SAM in the same system, it produces Cap 1 — reduced innate immunogenicity and higher translation efficiency.
Produced by E. coli recombinant expression with pharmaceutical-grade raw materials, and tightly controlled host-protein and nucleic-acid residuals.
Key Features
- Complete Cap 0 formation with a single enzyme: triphosphatase + guanylyltransferase + methyltransferase — no multi-enzyme assembly required
- Near-quantitative capping: ~100% within 1 hour, correct cap orientation
- Native cap structure: m7Gppp identical to the natural eukaryotic cap
- Cap 1 upgrade path: co-react with 2'-O-methyltransferase (SAM as methyl donor) in one system
- Clean residuals profile: endotoxin < 5 EU/mL, host protein <= 50 ppm, residual host DNA <= 100 pg/mg, nuclease / RNase controlled
Specifications
| Parameter | Specification |
|---|---|
| Composition | Recombinant D1 + D12 subunits, E. coli expression, pharmaceutical-grade raw materials |
| Activities | RNA triphosphatase, guanylyltransferase (GTP), guanine methyltransferase (SAM) |
| Capping efficiency | ~100% within 1 h, correct cap orientation |
| Activity definition | 1 U = the amount of enzyme that incorporates 10 pmol of GTP into an 80 nt transcript in 1 h at 37 C |
| Concentration | 10 U/µL |
| Pack sizes | 500 U / 10 KU / 100 KU / 500 KU / 1000 KU |
| Appearance | Clear liquid |
| pH | 7.5–8.5 |
| Purity | >= 95% |
| Bacterial endotoxin | < 5 EU/mL |
| Residual host DNA | <= 100 pg/mg |
| Residual host protein | <= 50 ppm |
| Nuclease / RNase residues | <= 10% substrate degradation in quality-control assays |
| Required cofactors | GTP (capping) and SAM (methylation) |
| Storage | -20 ± 5 C; aliquot on first thaw to avoid repeated freeze-thaw |
| Intended use | For Research Use Only |
Protocol Notes
- Pre-treat the RNA: IVT products must be purified first and dissolved in RNase-free water — not in EDTA or salt solutions.
- Resolve 5' secondary structure: heat the RNA at 65 C for ~5 min (up to 10 min for complex structures), then chill immediately on ice before starting the reaction.
- Prepare SAM fresh: dilute the 32 mM stock to a 2 mM working solution immediately before use — SAM is unstable at 37 C; keep it on ice.
- Cap 1 in one pot: run capping and 2'-O-methylation in the same system; an RNase inhibitor may be added.
- Re-purify the RNA after capping / polyadenylation before transfection.
Applications
- Enzymatic 5' capping of in vitro transcribed (IVT) mRNA — Cap 0 and Cap 1
- 5' end labeling of RNA
- RNA modification ahead of transfection or microinjection, improving stability and translation
- The upstream step of every mRNA-to-LNP workflow — cap your transcript first, then formulate with our Ionizable Lipids
Research Use Only
For Research Use Only. Not for direct administration to humans.
Ordering & Documentation
Pack sizes, cofactor supply options, COA, MSDS, ISO documentation, FOB pricing, MOQ and lead time are confirmed per inquiry: Send us an inquiry
| Product | Vaccinia Capping Enzyme (mRNA 5' Capping, Cap 0/Cap 1) |
|---|---|
| Catalog no. | VCE-500U/10KU/100KU/500KU/1000KU |
| Category | Lipids & Materials |
| Documentation | Certificate of analysis provided with every lot |
| Intended use | For Research Use Only (RUO). Not for diagnostic or therapeutic use. |
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