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物种
Human分子别名
rBoEnterokinase; Enteropeptidase; ENTK; PRSS7; hEK表达宿主
E.coli分子量
26 kDa (Reducing)
纯度
＞90% by SDS-PAGE标签
No Tag性状
Liquid缓冲体系
20mM Tris-HCl, 200 mM NaCl, 2mM CaCl2, 50% Glycerol, pH 7.4 @ 25°C
储存条件
Store at -25 ~ -15℃ for 2 years
文献引用
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.
2. Gasparian M E , Ostapchenko V G , Schulga A A ,et al.Expression, purification, and characterization of human enteropeptidase catalytic subunit in Escherichia coli[J].Protein Expression & Purification, 2003, 31(1):133-139.
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.
5 U/μl hEK, 20mM Tris-HCl, 200 mM NaCl, 2mM CaCl2, 50% Glycerol, pH 7.4 @ 25°C
Optimal incubation times and enzyme concentrations must be determined empirically for a particular substrate. Typical reaction conditions are as follows:
1、Combine 500 ug of sample with reaction buffer *（Recommended Reaction Buffer: 20 mM Tris-HCl, 50 mM NaCl, 2 mM CaCl2 (pH 8.0)）
2、Add 1 U of Enterokinase，
3、Incubate at 25°C for 16 hours
Enterokinase is inhibited by high salt concentrations. For optimal activity NaCl concentration should be 50mM or less. The pH of the buffer should be between 6 and 9. The enzyme requires 2 mM Calcium for activity.
- 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.
生物活性
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The substrate of 500μg fusion protein was digested by enzyme, and the addition amount of enterokinase in the sample was 0.2U, 0.5U, 1U,1.5U, 2U,2.5U and 3U respectively. The substrate is a fusion protein with a molecular weight of 30.5 KD and reacted at 25 ℃ for 16 hours, and the target band of 27.6kD is produced after restriction enzyme digestion.
电泳(SDS-PAGE)
2μg (R: reducing condition, N: non-reducing condition).
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