Dilution buffer reagent N9016

proteaseproteinase Kfor research

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 • 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 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. Preparation Instructions Stock solution can be prepared as 40-80mg/ml in dilution buffer [20 mM Tris-HCl (pH 7.4), 1 mM CaCl2] or [20 mM Tris-HCl (pH 7.4), 1 mM CaCl2, 2% Glycerol], sterilized using 0.22μm filter and supplied at final concentration of 20-40mg/ml in 50% 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.