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Proteinase K |
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Description:
Proteinase K is a non-specific, subtilisin-related serine protease with a very high specific activity. Proteinase K is not inactivated by chelating reagents such as EDTA or detergents such as SDS and is active over a wide range of pH (4-12.5).
Source:
Tritirachium album
Enzyme Properties
Specific Activity:
30 units/mg
Reaction & Storage Conditions
Unit Definition:
One unit is defined as the amount of enzyme required to liberate folin-positive amino acids and peptides corresponding to 1 µmol of tyrosine in 1 minute at 37°C in a total reaction volume of 250 µl (1).
Concentration:
20 mg/ml
Storage Conditions:
20 mM Tris-HCl
1 mM CaCl2
50% Glycerol
pH 7.4 @ 25°C
Storage Temperature:
-20°C
FAQs
- What are the guidelines for using Proteinase K?
- What is the Proteinase K activity in commonly used buffers?
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
16-Hour Incubation:
A 50 μl reaction containing 1 μg of λDNA (HindIII digest)
and 2 units of Proteinase K incubated for 16 hours at 37ºC
resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.
Exonuclease Activity:
Incubation of a 50 μl reaction containing 6 units of Proteinase K with 1 μg of a mixture of single and double-stranded
[3H] E. coli DNA (105 cpm/μg) for 4 hours at 37ºC
released < 0.2% of the total radioactivity.
Endonuclease Activity:
Incubation of a 50 μl reaction containing 2 units of Proteinase K with 1 μg of
ΦX174 DNA for 4 hours at 37ºC resulted
in < 5% conversion to RFII as determined by agarose gel electrophoresis.
RNase Assay:
Incubation of a 50 μl reaction containing 6 units of Proteinase K with 1
μg of MS2 RNA for 1 hour at 37ºC
resulted in no detectable degradation of the RNA as determined by agarose gel electrophoresis.
References
- Anson, M. L. (1939) J. Gen. Physiol., 22, 79.
Companion Products
Enterokinase, light chain
Factor Xa Protease
Furin
Genenase I
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