Vaccinia Capping Enzyme

Specifications:

Source: Vaccinia capping enzyme expressed in E. coli
Optimum Temperature: 37°C
Storage Buffer: 20mM Tris-HCl pH 8.0, 100 mM NaCl, 1 mM DTT, 0. 1 mM EDTA, 0.1% Triton X-100, 50% glycerol
Unit Definition: One unit is defined as the amount of enzyme needed to incorporate 10 pmol of (α-32P) GTP into an 80-nucleotide transcript within 1 hour at 37°C.

Overview:

In eukaryotes, mRNA undergoes post-transcriptional modification to form a specialized cap structure at the 5' end, which is essential for mRNA stability, transport, and translation. SignalChem Diagnostics Vaccinia Capping Enzyme catalyzes the formation of 5’ cap structure. The enzyme consists of two subunits, D1 and D12, and exhibits three enzymatic activities: RNA triphosphatase, guanylyltransferase, and guanine methyltransferase. It facilitates the attachment of a 7-methylguanosine (m7Gppp) cap to the 5' end of RNA (m7Gppp5'N).

In the presence of the appropriate concentrations of capping buffer, GTP, and S-adenosyl-methionine (SAM), Vaccinia Capping Enzyme can efficiently cap RNA within one hour while ensuring the correct orientation of cap structures.

This product is supplied as a sterile liquid and is suitable for in vitro and in vivo capping reactions of pre-mRNA, as well as 5'-end labeling of mRNA.

Components:

10X Capping Buffer: 150 μL (for 500 U); 500 μL (for 2,000 U); 1.25 mL x 2 (for 10,000 U)
Vaccinia Capping Enzyme (10 U/μL): 50 μL (for 500 U); 200 μL (for 2,000 U); 1 mL (for 10,000 U)

Storage and Shipping:

Store at -15°C to -25°C for up to one year. Ship on dry ice. Avoid repeated freeze-thaw cycles.