28 kDa (Reducing)
20 mM Tris-HCl, 200 mM NaCl, 2 mM CaCl2, pH 7.4 @ 25°C
Store at -25 ~ -15℃ for 2 years
1.Yong S Z, Yong C J, Yuan C X, et al. Secretory Expression, Purification and Characterization of Bovine Enterokinase Light Chain[J]. Journal of Nanjing University (Natural Sciences), 2004.
2.Haidong, Tan, and, et al. Purification and refolding optimization of recombinant bovine enterokinase light chain overexpressed in Escherichia coli[J]. Protein Expression & Purification, 2007.
Enterokinase (enteropeptidase, EC 3.4.4.8) occupies a key position in the utilization of dietary proteins. The enzyme initiates intraluminar digestion of proteins by the proteolytic conversion of trypsinogen to trypsin, which in turn activates the other pancreatic zymogens (Kunitz, 1939a,b; Hadorn et al., 1969). The proteolytic attack of enterokinase is directed exclusively toward the Lys6-Ile7 peptide bond of trypsinogen leaving all other lysine and arginine bonds in the molecule unaffected (Maroux et al., 1971). The resultant cleavage produces the simultaneous release of active trypsin and of the
amino-terminal hexapeptide Val-(Asp)d-Lys (Rovery et al., 1953; Davie and Neurath, 1955). This unique specificity exhibited by enterokinase is of interest as it relates to both the molecular basis of substrate recognition and the control of the digestive process.
5 U/μL Enterokinase, Bovine in 20 mM Tris-HCl (pH 7.4 @ 25°C), 200 mM NaCl, 2 mM CaCl2 and 50% (v/v) glycerol
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 light chain
3. Incubate at 25°C for 16 hours
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.
The results of 100μg substrate digestion separated under different quantity of UA070001, the reaction was incubated for 16 h at 25°C.
M marker
Lane 1 Only substrate
Lane 2 Substrate/ UA070001 = 4000/1 (1U)
Lane 3 Substrate/ UA070001 = 2000/1
Lane 4 Substrate/ UA070001 = 1000/1
2μg (R: reducing condition, N: non-reducing condition).
98%
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