Source: Yeast derived
Purity: >98% by SDS-PAGE
Endotoxin Level: N/O
Molecular Weight: 28.93kDa

Activity Assay

Proteinase K activity can be measured using hemoglobin as the substrate, >30U/mg

Description

Proteinase K is a serine proteas and belongs to Peptidase family S8. This enzyme has been widely exhibits substrate specificity and degrades many proteins in the native state. Proteinase K is inhibited by serine protease inhibitors such as phenylmethylsulfonyl fluoride. This enzyme activity is unaffected by the sulfhydryl modifying reagents. It is stable in EDTA buffer. 

Reconstitution

Briefly centrifuge the vial before opening. It is recommended to reconstitute the protein in sterile PBS containing 0.1% endotoxin-free recombinant human serum albumin.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles. In general: 12 months from date of receipt, -20 to -80°C as supplied. 1 month, 2 to 8°C under sterile conditions after reconstitution. 3 months, -20 to -80°C under sterile conditions after reconstitution.

How to make Stock Solution 

Proteinase K Stock Solution - 20 mg/ml (Download PDF format)

Instructions

1. Tris-CaCl₂ buffer PH8.0 (200mM Tris PH8.0+3mM CaCl₂)
   1. Measure 5 ml of 200mM Tris Buffer (pH 8.0), RNase free.
   2. Add 1.66 mg (3mM) calcium chloride (CaCl₂) to the Tris Buffer and dissolve. 
2. 5ml Stock Solution (20mg/ml)
   1. Measure 2.5 ml of the Tris-CaCl₂ Buffer (pH 8.0) into a centrifuge tube.
   2. Add 100 mg Proteinase K (YPK-K-100GM).
   3. Mix gently to dissolve the Proteinase K. 
   4. Sterilize the solution with a 0.22 μm filter. 
   5. Fill to a final volume of 5 ml with 100% glycerol (add 2.5ml glycerol). 
   6. Mix until completely dissolved. 

                 Store in aliquots at -20°C for 1 year. 

         3. Suggested Work Concentration: 0.05~1mg/ml

Note: Proteinase K interacts with nucleases. To ensure maximum Proteinase K activity, use water that has been treated with 0.1% DEPC