Hy-Proteinase K
Cat No. N9016

Size: 100mg
Contents: Hy-Proteinase K 100mg
Specific Activity: 40 U/mg protein
Form: Contains 100 mg Hy-Proteinase K, Lyophilized Powder


Description
Hy-Proteinase K is an endolytic protease that cleaves peptide bonds at the carboxylic sides of aliphatic, aromatic or hydrophobic amino acids. The Hy-Proteinase K is classified as a serine protease. The smallest peptide to be hydrolyzed by this enzyme is a tetrapeptide.

Features
Active in a wide range of reaction products

Applications
  • 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

Storage
For long time storage, store the Proteinase K powder at 4C and Delution Buffer at -20℃ separately. For use, dissolved solution should be stored at -20C.

Quality Control
DNase Activity: None detectable enzyme activity with λDNA after 6 hrs incubation at 37C.
RNase Activity: None detectable ribonuclease activity after 16 hrs incubation at 25C.


Definition of Activity Unit
One unit of the enzyme liberates Folin-positive amino acids and peptides corresponding to 1 μmol tyrosine in 1min at 37C, pH 7.5 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.

Source
The absence of ribonucleases is confirmed by appropriate quality test, functionally tested for digestion of template DNA after in vitro transcription.

Molecular Weight 
28.9 kDa monomer.

Dilution Buffer 
50mM Tris-HCl (pH 7.5), containing 5mM calcium chloride and 50% (v/v) glycerol.

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 95C for 10 minutes.