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                    Products > DNA Purification Accessary Reagents > Proteinase K (Lyophilized Powder)(N9016)

                    Proteinase K (Lyophilized Powder)(N9016)

                    Proteinase K

                    Catalog #

                    Size

                    Price

                    N9016

                    100mg


                    Contents:   Proteinase K     100mg
                                      

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

                    Features
                    Active in a wide range of reaction products

                    Description
                    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.

                    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 with λ DNA after 6 hrs incubation at 37oC.
                    RNase Activity: None detectable ribonuclease activity after 16 hrs incubation at 25oC. 

                    Source
                    Pichia pastoris cells with a cloned gene encoding Tritirachium album endolytic protease (Proteinase K).

                    Molecular Weight
                    28.9 kDa monomer (6).

                    Dilution Buffer
                    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.

                    Storage 
                    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. 

                    Note

                    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.


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