| <b>Unit Definition</b> One unit is defined as the amount of enzyme required to release 1.0 µmol of 4-nitroaniline per minute from N-Succinyl -Ala-Ala-Pro-Phe-p-nitroanilide at 25°C, in a total reaction volume of 105 μl. <b>Storage Buffer</b> 20 mM Tris-HCl 1 mM CaCl2 50% Glycerol pH 7.4 @ 25°C <b>Heat Inactivation</b> 55°C for 10 minutes <b>Molecular Weight</b> Apparent | 29 kDa <b>Unit Assay Conditions</b> A series of dilutions of Thermolabile Proteinase K are incubated with 0.25 mM N-Succinyl -Ala-Ala-Pro-Phe-p-nitroanilide in 0.1% SDS, 0.1% Triton X-100, 20 mM Tris-HCl, 5 mM CaCl 2 , 50 mM NaCl (pH 8.0 @ 25°C) in a 105 μl reaction. The reaction mix is incubated at 25°C. Liberation of p-nitroaniline is detected by real-time UV spectroscopy at 405 nm. |
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