AAT Bioquest

How do I dissolve proteinase K?

Posted September 7, 2023


There are several steps one should take to dissolve proteinase K properly. The steps below describe how to dissolve a proteinase K stock solution of 20 mg/mL.

  1. Measure out 5 mL of 200 mM Tris Buffer (pH 8.0), RNase free
  2. Add 1.66 mg (3mM) calcium chloride to the Tris Buffer and dissolve
  3. Measure out 500 µl of the Tris-CaCl2 buffer into a centrifuge tube 
  4. Add 20 mg proteinase K and mix gentle to dissolve
  5. Sterilize the solution with a 0.22  µl filter
  6. Fill to a final volume of 1 ml with 100% glycerol
  7. Mix completely until dissolved and store in aliquots at -20°C for one year

There are also alternate protocols with different procedures. For example, 20 mg/mL of proteinase K can also be dissolved in 50 mM Tris-HCL (pH 8), 1-5 mM calcium acetate and stored at 2-8 degrees celsius. The ideal buffer for proteinase K will vary from experiment to experiment.

Additional resources

Proteinase K


Amplite® Universal Fluorimetric Protease Activity Assay Kit *Green Fluorescence*