Size: 100 mg
Specific Activity: 40 U/mg protein
Form: Contains 100 mg Proteinase K, Lyophilized Powder
Active in a wide range of reaction products
Proteinase K is an endolytic protease that cleaves peptide bonds at the carboxylic sides of aliphatic, aromatic or hydrophobic amino acids. The Proteinase K is classified as a serine protease. The smallest peptide to be hydrolyzed by this enzyme is a tetrapeptide.
- Isolation of genomic DNA from cultured cells and tissues
- Removal of DNases and RNases when isolating DNA and RNA from tissues or cell lines
- Determination of enzyme localization
- Improving cloning efficiency of PCR products
DNase Activity: None detectable enzyme activity with λ DNA after 6 hrs incubation at 37ºC.
RNase Activity: None detectable ribonuclease activity after 16 hrs incubation at 25ºC.
Pichia pastoris cells with a cloned gene encoding Tritirachium album endolytic protease (Proteinase K).
28.9 kDa monomer
50 mM Tris-HCl (pH 7.5), containing 5 mM calcium chloride and 50% (v/v) glycerol.
Definition of Activity Unit:
One unit of the enzyme liberates Folin-positive amino acids and peptides corresponding to 1 µmol tyrosine in 1 min at 37°C using denatured hemoglobin as substrate.
Enzyme activity is assayed in the following mixture: 0.08 M potassium phosphate (pH 7.5), 5 M urea, 4 mM NaCl, 3 mM CaCl2 and 16.7 mg/ml hemoglobin.
For long time storage, store the Proteinase K powder at 4ºC and Delution Buffer at -20℃ separately. For use, dissolved solution should be stored at -20ºC.
Inhibition and Inactivation:
Inhibitors: Proteinase K is not inactivated by metal chelators, by thiol-reactive reagents or by specific trypsin and chymotrypsin inhibitors. Phenylmethylsulfonyl fluoride and diisopropyl phosphorofluoridate completely inhibit the enzyme.
Inactivated by heating at 95°C for 10 minutes.
Optimum activity at 50-55°C.
Rapid denaturation of enzyme occurs at temperatures above 65°C.
The recommended working concentration for Proteinase K is 0.05-1 mg/ml. The activity of the enzyme is stimulated by 0.2-1% SDS or by 1-4 M urea.
Ca2+ protects Proteinase K against autolysis, increases the thermal stability and has a regulatory function for the substrate binding site of Proteinase K.
Stable over a wide pH range: 4.0-12.5, optimum pH 7.5-8.0.
Proteinase K (Lyophilized Powder) - 100 mg
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