CATALOG NUMBER: C0220114
SIZE: 100 IU
PROTEIN INFO: E. coli derived bovine Enterokinase
NCBI ID: AAA16035
Enterokinase (EK) is synthesized by duodenum cells involving digestion. It cleaves the N terminal peptide of trypsinogen to produce active form trypsin. The specific cleavage site for EK is four aspartic acids Asp-Asp-Asp-Asp-Lys following by a lysine, not following by proline.
Recombinant Bovine Enterokinase (bEK) as the light chain is a single glycosylated polypeptide chain containing 200 amino acids. A fully biologically active form is prepared by America Diagnostics using its proprietary recombinant protein technology modified from Vozza (1996) protocol.
GSIVGGSDSREGAWPWVVALYFDDQQVCGASLVSRDWLVSAAHCVYGRNMEPSKWKAVLGLHMASNLTSP QIETRLIDQIVINPHYNKRRKNNDIAMMHLEMKVNYTDYIQPICLPEENQVFPPGRICSIAGWGALIYQG STADVLQEADVPLLSNEKCQQQMPEYNITENMVCAGYEAGGVDSCQGDSGGPLMCQENNRWLLAGVTSFG YQCALPNRPGVYARVPRFTEWIQSFLLEM
HOST: E. coli
FORMAT: Dry Powder
REACTIVITY: Unit Definition: One unit is defined as the amount of enzyme needed to cleave 50 μg of fusion protein in 16 hours to 95% completion at 22°C in a buffer containing 25mM Tris-HCl, pH 8.0.
FORMULATION AND PURITY
Recombinant protein over-expressed in E coli strain BL21, and affinity purification from inclusion body by Ni-NTA resin. The protein is > 97% pure, as determined by SDS-PAGE. The concentration is measured by Nanodrop.
Fig 1. bEK purified from E coli inclusion body. L, protein ladder. protein amount: 1, 30 ug; 2, 15 ug; 3, 7.5 ug.
STORAGE BUFFER: In PBS before lyophilized
RECONSTITUTION AND STORAGE
Upon received, save at freezer immediately if not used. After initial reconstitution with DI water (50-100uL), aliquot if necessary, and save the product at -80°C for future use.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to for technical protocols.
1. I. Yamashina. The action of enterokinase on trypsinogen. Acta Chem. Scand., 10 (1956), pp. 739-743.
2. Vozza, L. A., et al. (1996). "Production of a recombinant bovine enterokinase catalytic subunit in the methylotrophic yeast Pichia pastoris." Biotechnology (N Y) 14(1): 77-81.