The results of 100μg substrate digestion separated under diferent quantity of hEK-his, the reaction was incubated for 16 h at 25°C.
M marker
Lane 1 Substrate/ hEK-his = 500/1
Lane 2 Substrate/ hEK-his = 1000/1
Lane 3 Substrate/ hEK-his = 2000/1
Lane 4 Substrate/ hEK-his = 3000/1
Lane 5 Substrate/ hEK-his = 4000/1
Lane 6 Substrate/ hEK-his = 5000/1
Lane 7 Substrate/ hEK-his = 6000/1
(1 U hEK-his)
Lane 8 Substrate/ hEK-his = 8000/1
Lane 9 Substrate/ hEK-his = 10000/1
Lane 10 Only substrate
Recombinant Enterokinase, His tag (High specific activity)
Recombinant Enterokinase, His tag (High specific activity)
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$240 USD
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Product Details
Product Details
Product Specification
| Species | Human |
| Synonyms | Enteropeptidase, ENTK, hEK, hEK-His |
| Expression System | E.coli |
| Molecular Weight | 28 kDa (Reducing) |
| Purity | ＞95% by SDS-PAGE |
| Storage Buffer | 20 mM Tris-HCl, 200 mM NaCl, 2 mM CaCl2 (pH 7.4 @ 25°C) |
| Stability & Storage | Store at -25 ~ -15℃ for 2 years |
| Reference | 1.Melicherová, Kristína,Krahulec, Ján, afránek, Martin, et al. Optimization of the fermentation and downstream processes for human enterokinase production in Pichia pastoris[J]. Appl Microbiol Biotechnol, 2017, 101(5):1927-1934. |
Background
Enterokinase is a specific protease that cleaves after lysine at its cleavage site Asp-Asp-Asp-Asp-Lys. It will sometimes cleave at other basic residues, depending on the conformation of the protein substrate. This product is a high-purity, high-specific activity enterokinase prepared using a recombinant E.coli. It boasts a broad range of applicability (operating temperature: 4-45°C; pH range: 4.5-9.5) and retains partial activity in the presence of various detergents and denaturants.
Components
Storage Solution: 5 U/μL hEK-His in20 mM Tris-HCl, 200 mM NaCl, 2 mM CaCl2, pH 7.4 @ 25°C
Protocol
Optimal incubation times and enzyme concentrations must be determined empirically for a particular substrate. Typical reaction conditions are as follows:
Combine 500 ug of sample with reaction buffer
Recommended Reaction Buffer: 20 mM Tris-HCl, 50 mM NaCl, 2 mM CaCl2 (pH 8.0)
Add 1 U of Enterokinase light chain
Incubate at 25°C for 16 hours
Guidelines
1. Enterokinase is inhibited by high salt concentrations. For optimal activity NaCl concentration should be 50 mM or less. The pH of the buffer should be between 6 and 9. The enzyme requires 2 mM Calcium for activity.
Unit Definition
One unit is defined as the amount of enzyme required to cleave 500 µg of substrate to 95% completion in 16 hours at 25°C.
Picture
Picture
Bioactivity
SDS-PAGE
1μg (R: reducing condition, N: non-reducing condition).
SEC-HPLC
