Vaccinia Capping Enzyme ELISA Kit

In eukaryotes, mRNA is modified after transcription to form a special structure at the 5' end, namely the cap structure, which plays an important role in the stability, transport and translation of mRNA. Vaccinia virus capping enzyme is an effective enzyme that catalyzes the formation of the cap structure. It is composed of two subunits, D1 and D12, and has RNA triphosphatase activity, guanylyl transferase activity and guanine methyltransferase activity. It can connect the 7-methylguanine cap structure (m7Gppp) to the 5' end of RNA (m7Gppp5'N). Vaccinia virus capping enzyme, in the presence of appropriate concentrations of capping buffer, guanosine triphosphate (GTP), S-adenosylmethionine (SAM), etc., can cap RNA within one hour and ensure the correct direction.
This kit uses the experimental principle of double antibody sandwich enzyme-linked immunosorbent assay (ELISA) to detect the residual amount of vaccinia virus capping enzyme. Add the vaccinia virus capping enzyme standard and the sample to be tested to the ELISA plate (36709-A) pre-coated with anti-vaccinia virus capping enzyme antibody, and then add the diluted biotin-labeled vaccinia virus capping enzyme detection antibody (36709-C). Finally, add Streptavidin-HRP (SA-HRP) (36709-D) to form an antibody + antigen + antibody-Biotin + SA-HRP complex. After washing the plate, add TMB colorimetric solution (36709-H) for color development. TMB is converted from colorless to blue under the catalysis of HRP enzyme and finally converted to yellow under the action of stop solution (36709-I). The depth of yellow is positively correlated with the amount of vaccinia virus capping enzyme detected in the sample.

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