Proteinase K is classified as a non-specific serine protease. It is an endolytic protease that cleaves peptide bonds at the carboxylic sides of aliphatic, aromatic or hydrophobic amino acids.||Applications: |• Isolation of high molecular weight DNA |• Isolation of plasmid and genomic DNA |• Isolation of RNA |• Inactivation of RNase and DNase activities ||Advantages:|• Ready to use, provided in liquid form containing 40% glycerol.|• Smaller contamination risk with liquid form.|• Can be stored at -20°C still having the enzyme in liquid form.|• Excellent stability||Notes:|•Proteinase K is not inactivated by metal-chelating agents like EDTA, by thiol-reactive reagents (such as monoiodoacetic acid, p-chloromercuribenzoate) or by specific trypsin and chymotrypsin inhibitors. Phenylmethylsulfonyl fluoride and diisopropyl phosphorofluoridate completely inhibit the enzyme. |•Proteinase K is stable over a wide pH range: 4-12.5. |•Raising the temperature of the reaction from 37°C to 50°-65°C can increase the activity of Proteinase K.|•Proteinase K can be inactivated at 95°C for 10 minutes (some enzymatic activity may still remain)|•Proteinase K retains its activity in the presence of SDS (0.5-1%) or urea (1-4M).|•Depending on the application the enzyme is used in the absence or presence of 0.2-1% SDS or 1-4M urea.||Nomenclature: |EC=3.4.21.64; PROK; Endopeptidase K; Tritirachium Alkaline Proteinase||Source:|Tritirachium album limber||Concentration:|~20mg/ml||Form:|Supplied as a liquid in 10mM Tris HCl, pH 7.5, 1mM calcium acetate, 40% glycerol.||Recommended Dilutions: |The recommended working concentration for Proteinase K is 0.05-1mg/ml. Use 3ul of a 20mg/ml solution per 1.5ml of bacterial culture. Optimal dilutions to be determined by researcher.||Solubility: |Colorless to pale yellow, clear, complete||pH: |7.5 ± 0.2||Specific Activity: |≥30 units/mg||Unit Definition: |One unit liberates 1umole of Folin positive amino acids, measured as Tyrosine, at 37°C, pH 7.5, using urea-denatured hemoglobin as the substrate.||Quality Control:|Nicking Activity: None Detected|RNases: None Detected|Exonucleases: None Detected|Phosphatases: None Detected||Storage and Stability:|May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.