Enzyme reagent N9012

proteaseproteinase Kfor molecular biology

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. Features • Ready-to-use solution • Recombinant proteinase K • 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 Quality Control DNase Activity: None detectable enzyme activity after 6 hrs incubation with λ DNA at 37ºC. RNase Activity: None detectable ribonuclease activity after 16 hrs incubation with RNA at 25ºC. Source From yeast cells with cloned gene encoding genetically engineered Engyodontium album (Tritirachium album) endolytic protease. Molecular Weight 29.3 kDa monomer. 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 , 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. Storage Buffer The enzyme is supplied in: 50 mM Tris-HCl (pH 7.5), containing Ca2+ and glycerol as stabilizers. Inhibition and Inactivation Inhibitors: Proteinase K is not inactivated by metal chelators, by thiol-reactive reagents or by specific trypsin and chymotrypsin inhibitors.