Protocol for use with Enterokinase, light chain (P8070)
Optimal incubation times and enzyme concentrations must be determined empirically for a particular substrate. Typical reaction conditions are as follows:
- Combine 25ug of sample with reaction buffer and H2O (if necessary) to a total reaction volume of 20uL
* Recommended Reaction Buffer: 20 mM Tris-HCl, 50 mM NaCl, 2 mM CaCl2 (pH 8.0)
- Add 1uL of Enterokinase light chain
- Incubate at @ 25°C for 16 hours
Notes: Enterokinase is inhibited by high salt concentrations. For optimal activity NaCl concentration should be 100mM or less. The pH of the buffer should be between 6 and 9. The enzyme requires 2 mM Calcium for activity.