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 -20C 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 37C to 50-65C can increase the activity of Proteinase K. Proteinase K can be inactivated at 95C 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 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 37C, 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 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for 6 months after receipt at -20C.
Molecular Weight:
28.9
Form:
Supplied as a liquid in 10mM Tris HCl, pH 7.5, 1mM calcium acetate, 40% glycerol.
CAS Number:
[39450-01-6]
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