Enterokinase Cleavage Enzyme (EK1) is a highly pure and active endotoxin-free enterokinase that is produced using human cell line in serum free media. It is highly specific for cleaving fusion proteins with the recognition sequence, Asp-Asp-Asp-Asp-Lys (DDDDK). Because it is produced using human cell line, it has the appropriate glycosylation to catalyze highly accurate cleavage reaction with minimum non-specific activity. Its activity is stable over a broad temperature range, and it completes the reaction from 4°C to 60°C. |
Unlike other site-specific proteases that cut within the recognition sequences leaving extra amino acids in the cleaved peptide products, the EK1 cleaves after the C-terminal end of the lysine residue, and the fragment produced from the cleavage reaction does not inherit any residues from the DDDDK recognition sequence. Therefore, the application can be extremely advantageous for producing a 100% native protein sequence and structure from recombinant fusion protein, which has the desired product immediately after the enterokinase recognition sequence, DDDDK.
Owing to the presence of a His tag , EK1 can be easily removed after the cleavage reaction by affinity chromatography with Ni-IDA Agarose Beads. In addition, DDDDK is a part of the octapeptide FLAG tag (DYKDDDDK), which can be utilized as a fusion tag for recognition by antibody, and for detection of fusion protein expression with Western blot analysis, as well as for purification of the fusion protein by Anti-FLAG affinity chromatography. This array of applications makes EK1 an ideal tool in the research involving the study of protein structure and function, and protein production where native protein structures and sequences are desired.
Special Features
- Endotoxin free (i.e., produced in mammalian cells and purified from serum free media)
- lycosylated for improved efficiency and stability
- Rapid cleavage (95% cleavage in 1hr)
- Highly specific with minimum non-specific cleavage (even at 20x concentration)
- Stable over broad temperature range
- His-tagged for convenient removal post reaction