Enterokinase is a high-purity recombinant bovine enterokinase. the enzyme is purified by HPLC, with high purity, high specificity, and no other proteas. Enterokinase (EC 3.4.21.9) is a specific proteas that can cleave the carboxy-terminal site of lysine containing four aspartic acids in the front: Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid-Aspartic acid Aspartic acid-lysine. Enterokinase can remove the fusion protein located at the N-terminus of the protein to remove unwanted fusion tags.

High specificity
1) A specific proteas cleaves the carboxy-terminal site of lysine containing four aspartic acids in front: aspartic acid-aspartic acid-aspartic acid-aspartic acid-lysine (DDDDK)
2) No other protease, no non-specific cutting

Enzyme digestion conditions:
According to the definition of enzyme activity, an example of recombinant EK digestion conditions: In 25mM Tris-HCl 8.0 system:
Fusion protein concentration 0.1-1mg/ml (total protein 50-100g)
Reconstituted EK dosage 1-2U
Temperature 25C
Overnight digestion, or complete digestion requires 12h-16h.

Common factors affecting enterokinase activity
at >200mM Imi, or >200mMNaCl, or >5% glycerol, the digestion effect will be affected. You can refer to the following recommended methods for digestion:
1) in order to obtain an ideal digestion result, please dialyze the sample into 25mM Tris-HCl 8.0 buffer, and then perform digestion.
2) If it is inconvenient for dialysis, you can dilute the sample. the imi content is below 100mM, the NaCl concentration is below 50mM, and the glycerol concentration is below 5% for digestion, and the ratio of enzyme dosage to protein remains unchanged (ie 1U digestion 50μg protein).
3) If the sample solution contains one or more of the above components and it is inconvenient to remove, at this time, increase the enzyme amount or extend the digestion time to achieve better digestion results.